1

GENERAL GUIDELINES
ESTABLISHMENT OF
BIOSAFETY LEVEL-3
LABORATORY
INDIAN COUNCIL OF MEDICAL RESEARCH
DEPARTMENT OF HEALTH RESEARCH
MINISTRY OF HEALTH & FAMILY WELFARE
GOVERNMENT OF INDIA
NEW DELHI
 PREFACE
Globally, there is a strong perception that studies on microbial agents need more care and caution to
avoid health hazards to the laboratory personnel and the community. Emerging and newly emerging
diseases caused by unknown as well as known pathogens occur unpredictably worldwide. These often
appear in epidemic forms and are very difficult to diagnose and manage.

It is pertinent to have laboratories with high containment facilities (BSL-3) and protocols to deal with
such threats. The institutes with a mandate to investigate such outbreaks are required to be in a state
of preparedness. It is also mandatory that storage of these pathogens be done in appropriate biosafety
2 3
laboratories. Diseases of global public health significance are tackled by forming international networks
of laboratories having suitable infrastructure and trained manpower, and there is an increasing need
for containment facilities. The containment facilities are also required to enhance countries’ capacity on
preparedness in term of diagnostics and vaccines.

Biosafety policies and procedures are designed to safeguard personnel and the environment from
biologically hazardous materials and to comply with regulatory requirements. Containment science
emerged as an important branch of science in recent times. There were no national guidelines for
establishment, operation and maintenance, auditing and validation of containment laboratories.
Therefore, according highest priority to establishing such facilities becomes laboratory management’s
responsibility and lacunae in knowledge to execute such projects become biggest hurdles.

In most of the developing countries, the awareness about bio-containment has increased but planning,
designing, constructing and operating BSL-3 laboratories needs regular updates and clear definitions
of risk groups and their handling.

Realizing the importance of containment science these National Guidelines are developed to benefit
principal investigators, engineers, architects and management in conceptualizing the concept, layout
to final drawing, developing cost estimate to the execution of project. It has incorporated the hand-on
experiences of the contributors in establishing containment facilities in this country as well as in many
SEAR countries. This also has compiling the right information on a large number of areas that needs
tremendous attention during establishing such facilities.

Dr. Devendra T Mourya Dr. Nivedita Gupta Dr. Raman Gangakhedkar

ICMR Chair in Virology & Scientist F Div. of ECD Scientist ‘G’ & Head ECD
Zoonoses Indian Council of Medical Indian Council of Medical
Indian Council of Medical Research, New Delhi Research, New Delhi
Research, New Delhi
 FOREWORD
Biosafety and biosecurity are essential pillars of international health safety. In today’s interconnected
world, there is an ever-present threat of emerging, re-emerging and newly emerging viruses and
other pathogens that are capable of causing outbreaks & epidemics. Such emerging pathogens can
potentially and rapidly spread due to very fast air-transit and increasing urbanization in the era of
globalization. Many of these zoonotic viruses and other infectious etiologies are highly pathogenic and
transmissible in humans with high consequences.

High containment laboratories like Biosafety level-3 and -4 levels and specialized know-how for
4 5
handling these highly pathogenic micro-organisms and viruses are very essential in the developing
world, which faces the major brunt of infectious diseases and many of the outbreaks due to emerging
and re-emerging pathogens.

In Indian settings, recent outbreaks due to emerging viruses such as Nipah virus, arboviruses such
as Crimean-Congo hemorrhagic fever virus (CCHF) have led to increased emphasis on having high
containment facilities for the preparedness to handle outbreaks due to highly infectious pathogens.
Several high containment BSL-3 facilities have been established or currently in the planning or
execution phase. It is recognized that highly specialized expertise required for establishment,
maintenance and operation of high-containment facilities at BSL-3 and BSL-4 levels laboratories.

Currently, there is no existing guideline for establishing BSL-3 facilities for biomedical research and
diagnostic laboratories from the South Asian or South-East Asian region.

The Department of Health Research and the Indian Council of Medical Research has taken an initiative
to develop “General Guidelines for Establishment of Biosafety Level-3 Laboratories”.

The first edition of guidelines on establishment of BSL-3 laboratories will be a useful reference to
institutions in India and SEAR countries. It is hoped that these guidelines will act as a standardized set
of guidelines useful at the national level and the regional level for other countries in South-East Asia as
well as for other developing countries elsewhere.

Among many essential aspects covered in this guideline are clarifications on different biosafety
levels, objectives and scope of BSL-3 laboratory facilities, specific laboratory designs, considerations
in construction, staffing as well as pertinent issues related to operation and maintenance of these
facilities.

The guideline is timely and essential in the backdrop of outbreaks of emerging and re-emerging
highly-pathogenic viruses and micro-organisms such as Nipah virus, Avian influenza virus, SARS and
CCHF virus and drug-resistant mycobacteria in India and the South-East Asian region.

These guidelines will play an important role in the current efforts by the Government of India to
enhance biosafety and biosecurity in the country in accordance with the International Health
Regulations (IHR).

Prof. Balram Bhargava,

The Secretary, Department of Health Research, Govt. of India

& Director General, Indian Council of Medical Research
V. Ramlingaswami Bhawan, Ansari Nagar, New Delhi – 110 029
 ACKNOWLEDGEMENTS

6 7

We gratefully acknowledge the inputs from Lt. Gen D Raghunath, former Director General, Armed
Forces Medical Services for his valuable suggestions for the development of this guideline.

We sincerely acknowledge the experts from different areas, Dr. Ashwin Raut, NIHSAD, Indian Council
of Agricultural Research, Bhopal, Dr. Amit Singh, Indian Institute of Sciences, Bangalore, Dr. Arunava
Dasgupta, Central Drug Research Laboratory, Lucknow, Mr. Yogesh K. Shelar CDSCO, Ministry of Health,
Dr. Nitin K Jain, Department of Biotechnology, Dr. Umesh Gupta- Consultant, Indian Council of Medical
Research, for their opinion and suggestions for improvement of this document.

Contributors also sincerely thank the team of Indian Council of Medical Research; Dr. RR Gangakhedkar,
Dr. Nivedita Gupta, Dr. Rajni Kant and Department of Health Research; Ms. Anu Nagar, Dr. Ira Praharaj, Dr.
Neeraj Aggarwal, Dr. Harmanmeet Kaur, Dr. Jitender Narayan, Dr Neetu Vijay and Dr Siddharth Giri for
their valuable inputs for in the preparation of guideline.
 CONTENTS
Introduction 2
Identifying prerequisites for the construction 4
Defining basic objectives and scope of work 8
Preparation of pre-design 10
Preparation of BSL-3 laboratory design 12
8 1
Construction 16
Commissioning 20
Validation of the laboratory 24
Operation and maintenance 26
Further Reading 28
Annexure-1: BSL-3 Laboratory Infrastructure And Environment 30
Annexure-2A: Classification Of Infective Microorganisms By Risk Group 31
Annexure-2B: Relationship Of Risk Groups To Biosafety Levels, Practices And Equipment 31
Annexure-3: Points to be considered during Biological Risk Assessment 32
Annexure-4a: Criteria for BSL-3 laboratory 34
Annexure-4b: Criteria for BSL-3 laboratory 34
Annexure-4c: Criteria for BSL-3 laboratory 35
Annexure-5: Self-assessment of Biosafety 36
Annexure-6: Format for Biosafety Level 3 [Basic laboratory safety survey] 37
Annexure-7: BSL-3 Construction related information 42
Annexure-8: Conceptual BSL-3 Drawings 43
Table-1: Categories of laboratory and secondary barrier requirements 44
Table 2. Onsite equipment in the laboratory 47
Table-3: Thumb-Rule Calculations for Understanding Estimated Budget of Proposed 48
Laboratory
 INTRODUCTION
India, like many other progressing nations, faces a formidable challenge of
infectious diseases and especially of viral infections. Despite a few major
successes such as the elimination of wild polio virus, the country continues to
suffer substantial morbidity and mortality from both emerging and re-emerging
pathogens. The recent establishment of a 3-tiered network of Virus Research and
Diagnostic Laboratories (VRDL) across the country has greatly facilitated the timely
diagnosis of viral infections. These laboratories are equipped with facilities at the General considerations in the establishment of a Biosafety
Biosafety Level 2 (for handling pathogens causing moderate risk to humans and Level 3 (BSL-3) laboratories
2 low risk to the community), while a few of them have the capability to handle some 3
of the high-risk pathogens. Building preparedness against the emerging high-risk
An in-depth analysis of the scientific framework, objectives, pre-requisites (availability of
pathogens is a need of the hour in all parts of the country, and the establishment of
manpower, funding and space) as well as plan of proper utilization of the facility is a critical
more high containment laboratories would be a critical step to achieve this goal.
requirement prior to decisions on development of a biosafety level 3 laboratory.
A high containment Biosafety level 3 (BSL-3) laboratory would be mandatory for
all clinical, diagnostic, teaching & research facilities that perform work involving
agents that cause serious or potentially fatal disease in the workers through
Following essential steps are involved in establishing BSL-3
inhalation or lead to environmental contamination with these. All the work would laboratory (Figure-1)
be performed in bio-contained environments with appropriate engineering
controls. Work would be performed only by specifically trained personnel and
supervised by scientists competent in the handling of infectious agents.
Identifying pre-requisites for the construction

Defining basic objectives and scope of work

Preparation of pre-design, construction site selection

and risk assessment of area

Key elements of Biosafety level-3 (BSL-3) laboratory Preparation of BSL-3 laboratory design based on risk
assessment
Following are the key elements for the establishment of BSL-3 laboratory,

ŠŠ a) Biorisk assessment
ŠŠ b) Specialized physical, engineering infrastructure and environment (Annexure-1)
Construction of BSL-3 laboratory
ŠŠ c) Safety equipment (Primary and secondary containment barriers) and other supplies
ŠŠ d) Training of human resource for special practices
Commissioning of laboratory
Importance of these key elements are described and
discussed in relevant sections below:
The following guidelines are to summarize the practical approaches for the
Validation of the laboratory
establishment, commissioning, operationalization, requirement of validation and
functionality of a BSL-3 laboratory. These guidelines mainly assist the VRDL network
for establishing BSL-3 laboratories for expanding their diagnostic and research
capabilities in handling high risk pathogens. Operation and maintenance
 1. IDENTIFYING
PREREQUISITES
4
FOR THE 5

CONSTRUCTION
The first step in initiation for executing the thought of establishing BSL-3 laboratory is identifying the
pre-requisites/assessment of proposed facility for the construction is based on

ŠŠ Defined comprehensive outline of scientific framework

ŠŠ Specific objectives, pre-requisites (manpower, space and funding)
ŠŠ Plan for proper utilization of the facilities
ŠŠ Assessment of scientific and biosafety strength
ŠŠ Identifying the breach of biosafety with existing protocols
ŠŠ Preparation of preliminary flowchart for laboratory work
ŠŠ Specifying the need of engineering controls, Administrative controls and Personal protective equipment
(PPE) to accomplish the desired task.

Assessment of biorisk:
Based on the severity of infection caused by the pathogens to individuals and to the communities,
microbes are divided into four risk groups (Annexure-2a). Identifying the risk group of the pathogens
and other associated factors would be helpful in proper risk assessment.

Assessment of biorisk plays a key role in defining biosafety infrastructure, requirement of engineering
controls and safety equipment, hence in establishing BSL-3 laboratory.

Specifically, assessing the biorisk involved

ŠŠ In work protocols and high-risk pathogens to be handled.
ŠŠ With other factors: Pathogenicity, infectious dose, Natural route of infection, Potential outcome of
exposure, pathogen stability, availability of effective prophylaxis or therapeutic interventions.
ŠŠ During specimen handling, packaging and transport of high-risk pathogens in animal experiments to be
conducted
ŠŠ During discarding, disinfection and decontamination procedures
 ŠŠ Proposal for funding
Prerequisites for the construction of BSL-3/ ŠŠ Formulation of national
Assessment of proposed facility Committee to see the
development process Further, understanding the relationship of risk groups to biosafety levels, practices and equipment
ŠŠ Biosafety officer/In to be used (Annexure-2b) in laboratory procedures will help in understanding the requirements and
charge for facility
practices for risk reduction and containment of pathogens. Key steps involved in risk assessment are
represented in Figure-2 and important points to be considered during risk assessment process and risk
prioritization matrix is represented in Annexure-3.

Establishment of basic objectives Based on the above considerations a detailed conceptual proposal needs to be prepared including
ŠŠ Construction site budget estimation for funding, simultaneously formulation of national committee that will help in
selection and risk monitoring of developmental process.
assessment of area
ŠŠ Work flow chart
formation
ŠŠ List of Equipment
6 7
finalization
Predesign Identify hazards (agents
if known, lab procedures
and worker)

ŠŠ Hiring of contractor for

construction & Right
consultancy agency for
Design monitoring
ŠŠ Details of drawing
development & Evaluate/prioritize risks
evaluation by expert
ŠŠ Commissioning of onsite
equipment
ŠŠ Testing of all major
installations
ŠŠ Making SOPs for ŠŠ Monitoring of right
maintenance & manual for Construction construction
engineering ŠŠ Choosing right
ŠŠ Testing of all basic material as per the
resources commissioning & central public works Determine necessary controls
handing over of the facility department guidelines
ŠŠ Onsite equipment
ŠŠ Engineering controls
installation ŠŠ Administrative and Work
ŠŠ On time finishing of
Practice Controls
work
Commissioning
ŠŠ Personal Protective
ŠŠ Training program
equipment
preparation
Implement control measures
ŠŠ Validation of all major
equipment running &
SOP
ŠŠ Operation manuals, use & ŠŠ Emergency protocol
development of emergency
protocols
Validation development
ŠŠ Document preparation
ŠŠ Contingency plans for recording, manuals,
ŠŠ Drill of each SOP to validate SOPs of workflow,
& improve biosafety& primary &
ŠŠ Maintenance of equipment,
Evaluate effectiveness of
secondary barrier &
major installations testing
the control
ŠŠ Monitoring of human ŠŠ Evaluation from expert
health & work procedure & Operation and committee of each
data recording system & certification
maintenance
to use

Figure: -1 Essential steps involved in establishing BSL-3 laboratory Figure: -2 Risk Assessment process flowchart
Source: Mourya DT et al., (2014)
 2. DEFINING BASIC
OBJECTIVES
8
AND SCOPE OF 9

WORK

The basic objectives for establishment of containment laboratories for handling risk group-III agents
include:

ŠŠ Handling of clinical samples of patients infected with high risk pathogens during routine diagnostic
testing or outbreaks.
ŠŠ Conducting basic, applied or clinical research, or a combination of these on high risk pathogens.
ŠŠ Creating safe environment for detection, identification, propagation, manipulation of such organisms
in the laboratory; as well as maintaining safety of the community and the environment.
ŠŠ List of probable high-risk pathogens on which laboratory propose to work along with their relative
importance in terms of human morbidity and/or mortality in the region should be included in the
scope.
Further, considering the futuristic approach, scope of work intended to be performed in the BSL-3
laboratory has to be discussed thoroughly among the scientific/Biosafety officer and administrative
heads of the [Link], assessing the biorisk involved.

Defining basic
objectives and scope
of work
 During preparation of pre-design of the BSL-3 laboratories there are several criteria which play
critical role in pre design preparation that should be understood (Annexure-4a, b, c). Further knowing

3. PREPARATION different types of BSL-3 described below and their important features will ensure correct pre-design
development.

OF PRE-DESIGN Types of BSL-3 laboratories

Multiple designs of BSL-3 laboratories exist, varying in the overall layout, area, infrastructure and cost
input. In any format, these laboratories are sophisticated, expensive establishments, and the choice
of the design depends on several factors: the overall mandate, objectives and scientific framework
of the laboratory, the frequency of occurrence of high-risk pathogens in the region, clinical-scientific,
10 administrative and financial commitment, availability of expertise, laboratory space and funding 11
support. The features of the different types of BSL-3 laboratory and the factors to consider in their
choice are summarized in Table-1.

Selection of construction site and risk assessment of the area

ŠŠ BSL-3 laboratory should be ideally located in separate building or it must be separated from the
general traffic flow and accessed through an anteroom/air lock facility within a building.
ŠŠ Hence, for construction of BSL-3 laboratory a new stand-alone facility of the institutional site should
be selected. The construction site should meet the basic criteria of uninterrupted supply of water and
electricity, easy connectivity to the nearest airport and feasibility for a 3-tiered security system.
ŠŠ An in-depth risk analysis should be done in the area selected, for susceptibility to natural disasters like
landslides and earthquakes. The risk for possible damage to the site from occurrences like landslides,
heavy rains and flooding should be systematically evaluated.
ŠŠ The proposed site should be subjected to excavation to a depth of 2 meters to detect possible
presence of ‘borehole logs’. Detailed analysis of soil properties at the site also needs to be undertaken
in order to evaluate the suitability for construction.

Defining workflow for diagnostic/research procedures

Defining the work flow and preparation of flow charts based on mandate along with the utility of the
BSL-3 laboratory plays a primary role in enhancing the safety and effectiveness of development of pre-
design.

ŠŠ Identify the workflow, prepare required flow charts for each routine diagnostic and research
procedures.

Identifying the list of equipment

ŠŠ Equipment are the integral part of the BSL-3 laboratory for the conduct of experiments/ procedures.
ŠŠ Based on the workflow and the flow charts prepared for the conduct of tests, identify the equipment
required for specific work.
ŠŠ Finalize the list of equipment required and identify the rooms/space where these equipments should
be placed (Table-2).
ŠŠ Other equipment and accessories required for environment protection and personal protection
should also be listed for pre-design preparation i.e.: use of HEPA filtration of exhaust air, effluent
decontamination or chemical kill tank, personnel shower in the changing area, etc. inner and outer
change rooms for showers, to allow easy entry and exit protocols for the laboratory personnel.
 4. PREPARATION
OF BSL-3
12
LABORATORY 13

DESIGN

In this phase, complete design with action plan for constructing a BSL-3 laboratory should be
crystallized which mainly include:

Developing an action plan

A complete action plan for constructing the BSL-3 laboratory should be developed during the planning
phase. This involves preparing a detailed flowchart of the construction work and a schematic drawing
to enable detailed planning.

ŠŠ A detailed drawing of development of the entire facility should then be prepared, in order to monitor the
work progress.
ŠŠ The drawings must depict the layout of all laboratory areas to facilitate placement of essential on-site and
stand-alone equipment (including autoclaves, biological liquid effluent decontamination plant/chemical
kill tank, air handling units, exhaust filters).
ŠŠ The plan should also indicate the placement of safety equipment, such as fire extinguishers etc. within the
facility. A detailed concept proposal with detailed drawings, plans as per requirements and abiding the
biosafety regulations should be developed.
ŠŠ Preparation of detailed construction document with final specifications
ŠŠ Finalizing the engineering controls

Hiring of contractor/agency for construction

ŠŠ Hiring qualified, experienced architects, engineers to prepare design and construction of the project in
addition to finding the right construction agency/contractor.
ŠŠ Construction agency should have (i) the minimum average annual turnover during the last three financial
years (as per their audited balance sheets)- this ensures the ability of agency to complete the project. (ii)
Successful and timely completion of at least one similar project, which involved construction, testing,
 commissioning and validation of BSL-2 /or BSL-3 laboratory including civil works, electrical works, HVAC
works, Building Management System, door interlocks, access control system, primary barrier containment
equipment, decontamination system, etc., during the previous five years.
ŠŠ The ability of construction agency for designing and planning, correct evaluation of architectural layout
plans, men and material movement plans, zoning plans, specialized systems and services schemes, services
and utilities schemes, laboratory commissioning and validation protocols, laboratory security protocols,
integration of laboratory and equipment should be assessed.
ŠŠ The most important part is providing correct magnitude/valuation of the project and completion on time,
customer satisfaction, cost overrun, if any and litigation, if any.

14 Evaluation of design by expert committee 15

ŠŠ Constitution of a Project Implementation Committee

A Project Implementation Committee (PIC) should be constituted, with expert members from scientific,
technical or engineering & architecture background, holding an extensive experience in design and
commissioning of biosafety laboratories.
ŠŠ This committee shall have the overall accountability and authority for construction of the laboratory.
A crucial member of this committee would be the official who holds the responsibility for the overall
functioning and safety of the laboratory.
ŠŠ The committee would recognize the amount of work involved before laboratory design is -initiated.
ŠŠ In view of the scarcity of advanced biosafety expertise in the country, a few members with scientific and
engineering background could be trained in currently existing BSL-3 laboratories, to gain exposure and
experience.
ŠŠ All members of the PIC should gain familiarity with the information relevant to the project. The team
should meet at least once every month, for discussions to evolve the criteria for the proposed laboratory,
based on priorities and requirements.
ŠŠ An evaluation of the prepared drawings should be done by an expert committee to review whether the
proposed design is as per the requirements and also complies with the “General Guidelines of Biosafety
and Biosecurity” for the BSL-3 laboratories in state/country.
ŠŠ Design should also consider placing vibration-sensitive instruments such as microscopes etc. away from
structural columns or over slab-on-grade.
ŠŠ The engineering staff associated with the laboratory should gain experience in modern mechanical
systems, including Building Management Systems that would be installed in the laboratory.
ŠŠ Design should also include those for electrical supply of transient or voltage fluctuations with harmonics
along with adequate voltage requirement of uninterrupted power supply (UPS).
ŠŠ Identification and appointment of staff who would perform specific responsibilities in the laboratory needs
to be done at this stage. Preliminary finish schedules and material selection for hardware and construction
requirements should be finalized. The recommendations also include developing an Engineering Manual
for the operation of the facility.
Defining all the above factors will help in monitoring the progress of the work and successful, timely
completion of the project.
 5. CONSTRUCTION

16 17

An important aspect in the construction of the BSL-3 laboratory is putting forward the conceptual
proposal with detailed drawings and plans, justifying the objectives for the construction as per the
requirements along with abiding with the national and international guidelines of biosafety.

ŠŠ A construction document with final specifications is to be prepared at this stage. The final proposal with
detailed drawings and equipment specifications should submitted for release of funds so that tenders can
be invited.
ŠŠ Tender must be prepared to select agencies/organizations experienced in similar works earlier, and with
proven track record.
ŠŠ The tender should include the general conditions of contract, description and scope of work, qualification
criteria, instructions to bidders and evaluation of bids, in addition to the notice inviting the tender.
ŠŠ Some of the construction documents setting basic criteria and requirements for this facility should be part
of the tender document.
ŠŠ A credible agency and contractor with successful track record in undertaking similar works should be
identified as described in laboratory designing phase.
ŠŠ Awarding the tender to agency, hiring a consultant agency for supervising the quality of construction as
per the requirements recommended in national and international standards.
ŠŠ Tender must be customized to select a professional organization, having experience of constructing this
kind of laboratory.
ŠŠ An agreement should be signed specifying all the requirements and guidelines to be followed, mentioning
the time limit given for the completion of the project and the penalty clause, if not completed in time.
 Monitoring of precise construction
ŠŠ It is important to monitor precise construction by Identifying project management consultant.
ŠŠ Given the scarcity of regulations, codes and standards for high containment laboratories in the country,
and the complexity of the international standards, a competent agency or group of engineers should
18 supervise the quality of construction process. 19

ŠŠ The local engineering department may ensure supervision of construction. Installation of integral
equipment and procurement of accessory equipment required for the laboratory could proceed along with
the construction work.
ŠŠ The team members should possess a thorough understanding of the features of the equipment being
procured and the essential requirements for their installation, calibration and routine maintenance.
ŠŠ Selection of correct material as per international guidelines is the prime requirement during construction
phase.

Installation and testing of equipment

ŠŠ The placement and installation of all on-site equipment like autoclaves, Biological Liquid Effluent
Decontamination (BLED) plant/chemical kill tank, air handling units, exhaust filters within the facility must
be identified.
ŠŠ Placement of all safety equipment like fire extinguishers etc. within the facility should also be incorporated
in the detailed drawing.
ŠŠ Approvals should also be taken at this stage from the local fire departments, so that at later stages facility
can pass the fire norms. Complete Heating Ventilation Air Conditioning (HVAC) design calculations for
maintenance of unidirectional airflow and negative pressure as compared to the ambient within the
facility and air flow diagrams must be prepared.
ŠŠ Adequate considerations must be given to the ability of the laboratory facility engineering staff to start
learning to operate modern mechanical systems as soon as these are installed in the laboratory.
ŠŠ During planning with various laboratory work flowcharts, on-site and stand-alone equipment and the
requirement of electrical supply of transient or low voltage fluctuations with low harmonics along with
adequate voltage requirement of uninterrupted power supply (UPS), should be taken care of.
ŠŠ At this stage, staff that will be required for working in the facility must be identified and trained for carrying
out specific responsibilities.
ŠŠ Developing an “Engineering Manual” for the operation of the facility is also recommended.
 6. COMMISSIONING

20 21

The process of laboratory commissioning plans ideally should start in the design phase and continue
throughout the construction process.

The commissioning process of the laboratory includes three phases:

(i) Testing and commissioning of “on-site” equipment– this should be initially performed by the construction
contractor alone. It should be repeated and demonstrated to an authorized person or project management
consultant for the facility.

(ii) Testing and validation of the commissioning process of equipment are performed in presence of the
facility In-charge and Biosafety Officer.

(iii) Final testing and commissioning should take place in presence of committee / project team. On
completion the laboratory is to be made functional, ready for takeover.

ŠŠ Commissioning procedure for the laboratory should be well designed and implemented to verify the safe
facility operation.
ŠŠ Testing and commissioning of some of the elements are crucial to the proper functioning of the
containment, such as airflow patterns and pressures within isolators and biosafety cabinets, temperature
profiles in autoclaves, procedures for decontamination and sterilization, verification of light lux level (must
be between 300-600 lux), operation of HVAC systems, capacity calculations of HVAC systems plant, chilled
water pumps capacity, air quantities at outlet diffusers / grilles, and air compressor, testing air curtains,
steam boiler, clean room garment storage cabinet, floor traps, drains, dunk tanks, checking of ceiling
panels, pass box, shower cabinets/air shower, water outlets, air leak in ducts as well as plenums, doors and
view panels along with functioning of all the alarm systems.
ŠŠ Taking over the facility includes verifying all the basic requirements as per the approved layouts, electrical
connections (raw, essential and UPS), local area network (LAN) connections, servers, water connections,
sewage connection, hardware fitting, telephones and intercoms, functioning of the BMS with all the
desired parameters, fine setting of access control and all the inventories.
 Staff Pattern in a BSL-3 Laboratory and Preparation of Training
program
ŠŠ A multi-disciplinary team of specially trained staff, consisting of clinical/scientific, engineering,
administrative and support staff is integral to the functioning of a BSL-3 laboratory.
22 23
ŠŠ An experienced and qualified Biosafety Officer and a Scientist-in-Charge would be responsible for
the overall functioning of the facility. The objectives of the laboratory and the specific projects to be
undertaken in the laboratory would be important considerations in deciding the pattern of scientific and
technical team.
ŠŠ Staff requirement would also include an officer-in-charge for maintenance, one technician each for HVAC,
electrical systems, instrumentation and the staff for general building and service maintenance.
ŠŠ On-site training on relevant aspects could be considered for the project staff, at other laboratories.

Preparation of Training program

All personnel recruited in the BSL-3 laboratory must successfully complete a comprehensive training
program on BSL-3 laboratory functioning.

ŠŠ This training program should be developed keeping in mind the mandate and specific requirements of the
laboratory; this training should cover the concepts of biosafety and biosecurity and safe working within
the BSL-3 laboratory.
ŠŠ The Biosafety Officer and Scientist-in-charge of the facility shall be responsible for the development and
implementation of the training programs and protocols.
ŠŠ In addition to the initial training, refresher trainings on biosafety (along with safety drills) should be
conducted every six months.
ŠŠ Records of orientation and refresher training for staff should be maintained along with the list of trainees
and results of assessment/competency evaluation.

Development of Standard Operating Protocols (SOPs)

ŠŠ SOPs should be developed for use of all laboratory equipment, general laboratory work, processing of
clinical samples and diagnostic assays performed.
ŠŠ Development and validation of SOPs for emergency protocols in the laboratory is another important
concern, and should be considered from the beginning of laboratory construction.
ŠŠ The SOPs developed should also be validated by a designated officer.

Testing of all resources and handing over of laboratory

After completing commissioning process all resources including SOPs should be tested before taking
over the laboratory from the construction agency to ensure and understand practical functionality as
well as user acquaintance of the BSL-3 facility.
 7. VALIDATION
OF THE
24 LABORATORY 25

Approval from local statutory authorities like the Fire Department and Municipal Corporation should
be obtained before commencing operation of the laboratory for validation. Staff training should be
reinforced and mock drills along with validation of SOPs should be conducted at this stage. Upon
completion of these processes, the validation process for the laboratory can be started.

Preparation of essential documents and program for laboratory

validation
Important step at this phase would be the preparation of documents certifying compliance with the
international guidelines. A document describing the mandate and features of the laboratory would be
a primary requirement.

ŠŠ In addition, commissioning reports of major equipment, SOPs of laboratory workflow, use of equipment
and engineering controls (including records of operation of AHU and change of filters), user log books of
all equipment, certification details, maintenance reports should also be prepared and maintained in the
laboratory.
ŠŠ The records for entry/exit, printouts of shower entry, records of daily checks, requisition file, and file of
calibration of equipment should be prepared and maintained.
ŠŠ Documents in support of validation of decontamination processes (e.g., spore strip test for validation
of autoclaves, records of area fumigation and surface swab tests), record of performance of Building
Management System, room pressure, temperature and humidity should also be prepared.
ŠŠ Documents related to the inventory of samples and other material (stored in laboratory refrigerators,
-20˚C and -86˚C freezers and liquid nitrogen storage) and backup plans should be developed.
ŠŠ Facility and Operation Manuals explaining biosafety aspects as well as maintenance of engineering
systems should also be prepared.
ŠŠ A Technical Manual should also be developed for the facility.
ŠŠ Validation of the laboratory should be conducted by internal and external expert committees in liaison
with the Facility-in-Charge and/or Biosafety Officer, construction agency and project management
consultancy.
ŠŠ The validation process aims to ensure biosafety and biosecurity concerns to the workers, work and the
environment as well as adherence to the work flow program.
 8. OPERATION AND
MAINTENANCE
26 27

The laboratory would commence the safe operation mode upon completion of the validation process
and obtaining the certificate to use the facility.

ŠŠ The contingency plan for emergencies needs to be reviewed during the regular operation mode, to prevent
biosafety breaches with respect to the building management systems, UPS, DG set and autoclaves.
ŠŠ All organizations which undertake work involving microorganisms/genetically engineered organisms
should constitute an Institutional Biosafety Committee which should prepare an updated site emergency
plan as per recommendations given in the Review Committee on Genetic Manipulation (RCGM), and also
evaluate the biosafety concerns arising from experimentation and containment issues.
ŠŠ Operation and comprehensive maintenance contract for day to day (24Hr) operation of BSL-3 lab should
be undertaken in advance with the suppliers for all major equipment to ensure prompt service. A contract
should also be undertaken for maintenance of the facility with the facility contractor to ensure engineering
support.
ŠŠ Finally, Memoranda of Understanding should be executed with the contractor and subcontractors to
ensure uninterrupted provision of spares and services, for a minimum period of 5 years.
After successful competition of all the phases involved in establishment of BSL-3 laboratory, during
routine usage of the facility, the laboratory persons have to self- assess the biosafety parameters
(Annexure-5) in the laboratory. Further to generate systematic documented evidence, every BSL-3
laboratory should develop a format for capturing basic laboratory safety check points (Annexure-6).

In conclusion, for achieving biosafety and biosecurity during handling high risk pathogens,
multifactorial assessment of biorisk, infrastructure, safety equipment, Good Laboratory Practices,
trainings etc. is mandatory as described in these guidelines.
28 29
FURTHER READING

ŠŠ Mourya DT, Yadav PD, Majumdar TD, Chauhan DS, Katoch VM. Establishment of Biosafety Level-3 (BSL-3)
Laboratory: Important criteria to consider while designing, constructing, commissioning & operating the
facility in Indian setting. Indian J Med Res 2014; 140:171-83.
ŠŠ Mourya DT, Yadav PD, Khare A, Khan AH. Certification & validation of biosafety level-2 & biosafety level-3
laboratories in Indian settings & common issues. Indian J Med Res 2017; 146:459-67.
ŠŠ Oregon State University. Basic Laboratory Design for Biosafety Level 3 Laboratories. Available at https://
[Link]/cpd-standards/appendix/room-and-space-types/basic-laboratory-design-biosafety-
level-3-laboratories.
ŠŠ A.N. Zaki, Biosafety and biosecurity measures: management of biosafety level 3 facilities. International
Journal of Antimicrobial Agents 36S (2010) S70–S74
ŠŠ World Health Organization. Laboratory biosafety manual 3rd edition (2004), World Health Organization,
Geneva, 2004.
ŠŠ Regulations and guidelines for recombinant DNA research and bio contentment 2017. Accessible at: http://
[Link]
 Annexure-1: BSL-3 Laboratory Infrastructure And Environment Annexure-2A: Classification Of Infective Microorganisms By Risk
Group
Sr.# BSL-3 laboratory infrastructure and environment Remarks
Risk Group 1 (No or A microorganism that is unlikely to cause human or animal disease
1 Controlled access
low individual and
2 Physical separation from access corridor community risk)
3 Personnel shower Optional, as/risk Risk Group 2 A pathogen that can cause human or animal disease but is unlikely to be
assessment (Moderate individual a serious hazard to laboratory workers, the community, livestock or the
4 Anteroom; two self-closing interlocked doors risk, low community environment.
5 Single pass air directional air flow risk) Laboratory exposures may cause serious infection, but effective treatment
and preventive measures are available and the risk of spread of infection is
6 Air pressure differential (ranging from -10 to -60 Pa, as per the
30 limited. 31
risk assessment)
Risk Group 3 (High A pathogen that usually causes serious human or animal disease but does
7 Exhaust system independent from remainder of the building
individual risk, low not ordinarily spread from one infected individual to [Link]
8 Supply system independent from remainder building community risk) treatment and preventive measures are available.
9 Single HEPA filtered exhaust Depending upon the size Risk Group 4 (High A pathogen that usually causes serious human or animal disease and
of the lab., multiple HEPA individual and that can be readily transmitted from one individual to another, directly
filters at exhaust points are community risk) or indirectly. Effective treatment and preventive measures are not usually
necessary. Thus, HEPA SAFE available.
CHANGE BOX could be
considered connected with
a single suitable blower
with 100% redundancy
10 Supply exhaust fans interlocked with supply fans
11 Redundant exhaust fan (N+1)
Annexure-2B: Relationship Of Risk Groups To Biosafety Levels,
12 Utilities backflow prevention
Practices And Equipment
13 Minimum 6-12 air changes per hour
14 Autoclaves available in facility Within facility preferable Risk Biosafety Laboratory Type Laboratory practices Safety equipment
15 Sealed windows Group level
16 Sealed penetrations 1 Basic Basic teaching, Good Microbiological None: open bench work
17 Seamless floors Biosafety research lab Techniques (GMT)
level 1
18 Monolithic ceiling
2 Basic Primary health, GMT plus protective Open bench plus biological
19 Chemical resistant floors, walls, ceiling, doors and frames clothing, biohazard safety cabinet (BSC) for
20 Chemical resistant piping, fixtures and casework Biosafety services; sign potential aerosols
21 BSL-3 laboratory on emergency power level 2 diagnostic,
services, research
22 Laboratory lighting and biosafety cabinets on emergency Preferred to avoid blackout
Inverter/UPS 3 Containment Special diagnostic As level 2 plus special BSC & other primary devices
Biosafety services, research clothing, controlled for all activities
23 HVAC failure alarm level 3 access, directional
24 Pressure differential monitors airflow
25 Deep sealed floor traps 4 Maximum Dangerous As Level 3 plus airlock Class III BSC, or positive
26 Hand wash sinks and emergency eye wash Containment pathogen units entry, shower exit, pressure suits in conjunction
27 Surface mounted sealed lighting Biosafety special waste disposal with Class II BSCs, double
level 4 ended Autoclave (through
28 Less than 60 db. noise level
the wall),
29 Lighting (300-600 lux)
30 BSC shall be located out of the mainstream of traffic filtered air
31 Isolation dampers in ventilation system
 Annexure-3: Other factors that should be considered as appropriate include:
Points To Be Considered During Biological Risk Assessment ŠŠ Pathogenicity of the agent and infectious dose
ŠŠ The backbone of the practice of biosafety is risk assessment. ŠŠ Potential outcome of exposure
ŠŠ While there are many tools available to assist in the assessment of risk for a given procedure or experiment, ŠŠ Natural route of infection
the most important component is professional judgment. ŠŠ Other routes of infection, resulting from laboratory manipulations (parenteral, airborne, ingestion)
ŠŠ Risk assessments should be performed by the individuals most familiar with the specific characteristics of ŠŠ Stability of the agent in the environment
the organisms being considered for use, the equipment and procedures to be employed, animal models
ŠŠ Concentration of the agent and volume of concentrated material to be manipulated
that may be used, and the containment equipment and facilities available.
ŠŠ Presence of a suitable host (human or animal)
ŠŠ The laboratory director or principal investigator is responsible for ensuring that adequate and timely risk
32 ŠŠ Information available from animal studies and reports of laboratory-acquired infections or clinical reports 33
assessments are performed, and for working closely with the institution’s safety committee and biosafety
personnel to ensure that appropriate equipment and facilities are available to support the work being ŠŠ Laboratory activity planned (sonication, aerosolization, centrifugation, etc.)
considered. ŠŠ Any genetic manipulation of the organism that may extend the host range of the agent or alter the agent’s
ŠŠ Once performed, risk assessments should be reviewed routinely and revised when necessary, taking into sensitivity to known, effective treatment regimens
consideration the acquisition of new data having a bearing on the degree of risk and other relevant new ŠŠ Local availability of effective prophylaxis or therapeutic interventions.
information from the scientific literature.
ŠŠ On the basis of the information ascertained during the risk assessment, a biosafety level can be assigned
ŠŠ One of the most helpful tools available for performing a microbiological risk assessment is the listing of to the planned work, appropriate personal protective equipment selected, and standard operating
risk groups for microbiological agents. However, simple reference to the risk grouping for a particular procedures (SOPs) incorporating other safety interventions developed to ensure the safest possible
agent is insufficient in the conduct of a risk assessment. conduct of the work.
ŠŠ No one standard approach or correct method exists for conducting a risk assessment. ŠŠ Each laboratory should adopt a safety or operations manual that identifies known and potential hazards,
ŠŠ However, several strategies are available, such as using a risk prioritization matrix, conducting a job hazard and specifies practices and procedures to eliminate or minimize such hazards.
analysis; or listing potential scenarios of problems during a procedure, task, or activity.

Risk Prioritization Matrix

ŠŠ The process involves the following five steps:
ŠŠ Identify the hazards associated with an infectious agent or material.
ŠŠ Identify the activities that might cause exposure to the agent or material.
ŠŠ Consider the competencies and experience of laboratory personnel.
ŠŠ Evaluate and prioritize risks (evaluate the likelihood that an exposure would cause a laboratory-acquired
infection and the severity of consequences if such an infection occurs).
ŠŠ Develop, implement, and evaluate controls to minimize the risk for exposure.
 Annexure-4a: Criteria for BSL-3 laboratory List of equipment on the emergency generator (specifications) AHU with twin
motors, Access
Overall guidelines High control, Isolators,
Containment Computers &
Laboratory LAN system,
(HCL) BSCs, Emergency
Containment barrier outlined and appropriate Yes lights and other
important
Office area located outside of laboratory Equipped with computer control for Yes
equipment
restricted access
are backed by
Clean/Dirty change area separated by a walk-through shower Yes inverter & UPS.
34 Double door pass-through autoclave with interlocking doors or visual/audible Yes Whole electrical 35
alarm system has back-
Large door to allow entry of equipment Yes up to back-up DG
sets.
Interlocking door system (Computer control access with manual override) Yes
Emergency lighting to be provided Supported by
Room pressure to be monitored either by differential pressure gauge or other Yes
Inverters and UPS
means of display of pressure.
Decontamination ports, Dunk tank, Pass through Yes
Surfaces having Epoxy and Polyurethane coating, Able to withstand Yes
disinfectants, No porous joints
Annexure-4c: Criteria for BSL-3 laboratory
Impact resistant material Yes
All penetrations are sealed (water plumbing/ducts/cables etc.) Yes HEPA Housings High
Air Handling separate from other areas of lower/non containment, Motorized Yes Containment
bubble tight damper or HEPA filter for backdraft protection. Bubble tight Laboratory
damper for gaseous decontamination. Exhaust completely sealed and ducked (HCL)
out Provided with a bubble tight damper for isolation on inlet and outlets Yes
Location of supply does not interfere with inlets and outlets; different users Yes Provided with fumigation ports upstream and downstream Yes
properly designed hence not applicable Provided with upstream injection port and downstream access port for Yes
All inlets and outlets are computerized to control and balance the negative Yes scanning purposes
pressure Leak tight Yes
Biosafety cabinets (BSCs)
Type and class of BSCs Class-II, B2 type
(thimble, hard-ducted, recirculated, charcoal filter, etc.) with HEPA in and
Annexure-4b: Criteria for BSL-3 laboratory HEPA out filters.
100% exhaust.
Laboratory Services High Chemical
Containment resistant
Laboratory (high density
(HCL) polyurethane
Communication system (Direct telephone line, Intercom, LAN network Yes material) used for
connections and CCTV etc.) ducting
Water supply provided with backflow preventers at containment barrier Yes Located 1.5m from supply/exhaust ducts, doors, air generating equipment Exhaust
Full-fledged shower facilities Yes completely
sealed and
Drainage traps with double U traps and filter mechanism Yes
ducted out
Effluent piping to be heat and chemical resistant NA
30 cm clearance between exhaust outlet and ceiling Yes
Autoclave condensate drain located on dirty side (leading to either BLED tank Autoclave
30 cm around cabinet for access Yes
or Chemical Kill Tank) condensate drain
located on dirty
side
 Annexure-5: Self-assessment of Biosafety Lab personnel receive appropriate immunizations or All the persons immunized with KFD killed
tests for agents handled vaccine, other immunizations not in the
Self-assessment of Biosafety institute’s general policy
ŠŠ Proper engineering controls are being used and are functioning adequately as designed
Insect and rodent control program available Yes, in place
ŠŠ Appropriate site and protocol specific administrative controls are in place
Eating or drinking and the storage of food are Yes, have defined area for eating and
ŠŠ Personal protective equipment is appropriate for the tasks being performed prohibited drinking
ŠŠ Decontamination of waste and materials has been adequately considered and proper waste Mechanical pipetting devices are used Yes, rubber bulb or battery-operated device
management procedures are in place Cultures and stocks decontaminated before disposal Yes, autoclaved before disposal
ŠŠ Proper procedures for general laboratory safety, including physical, electrical and chemical safety Plasticware substituted for glassware when possible Yes, mostly disposable plasticware are used
are in place.
Used needles, syringes and other sharps placed in Used needles should be destroyed using
36 Administrative Controls and Documentation All the manual, protocols & other material appropriate sharps containers needle cutter before discarding it in the 37
available to lab persons sharps container
Authorization for protocols on file Certain restricted protocols & other Pasteur pipettes properly disposed Disposable plastic pipettes used
confidential material available to specified
Broken glass is not handled directly but by Yes
persons on password digitized files
mechanical means
Biosafety Manual and SOPs Biosafety Manual and SOPs are available to
Work surfaces are decontaminated daily and Yes
lab persons
following spills
Emergency Response Plan including spill Present in the lab, available to lab persons
All bio-waste is properly disposed Yes, as per the corporation norms for
contamination
disposal of bio-waste
Documented Biosafety training for all personnel Present in the lab, available to lab persons
Laboratory Facilities
Biosafety Sign posted on entrance Yes
The laboratory has a sink & soap for hand washing Yes
Emergency shower/eyewash facility is available Yes Annexure-6: Format for Biosafety Level 3 [Basic laboratory safety
Windows that open to the outdoors are fixed w/ Windows in the main lab not open able, survey]
screens others with grill
First aid kit readily available Present in the lab, available to lab persons Location: ________________ Date: _____________
Safety Equipment – Primary Barriers Person in charge of laboratory:
Biological safety cabinets (BSCs) are used whenever Yes, have separate BSC for clean & infected Laboratory Yes No N/A
procedures with potential for creating infectious work. Proper signage: ultraviolet light, laser, radioactive material, etc.
aerosols or splashes are conducted
Appropriate biosafety guidelines available and followed
BSCs located away from doors, heavily traveled areas, Yes, location is secured place
Laboratory equipment properly labelled (biohazardous, radioactive,
etc. to maintain air flow
toxic, etc.)
BSC has been certified within the past year Yes, under AMC and checked twice in a year
Laboratory design
Centrifuge safety containment cups or sealed rotors Yes, also separate centrifuge for clean &
Designed for easy cleaning
with O-rings available infected work
Room ultraviolet lights on interlock switch
Lab coats are not taken home by lab personnel Yes, not to be taken out of working area
All shelves secured
Autoclave
Bench-tops waterproof and resistant to acids, alkali, organic solvents
Self-assessment of Biosafety
and heat
Is autoclave QC log kept Yes, the chemical indicator always used
Adequate illumination provided
Appropriate autoclave bags are used Yes, autoclavable bags are used
Adequate storage space available and appropriately used
The autoclave is monitored with biological Yes, once in a year
Gas cylinders (Should be housed outside BSL-3 core area, tubings
indicators
should be in embedded condition)
Standard Microbiological Practices
All cylinders secured
Access to laboratory is restricted Yes
Caps on reserve cylinders
Containers properly labeled Yes, for discarding infectious material,
broken glasses, waste gels etc.
 Asphyxiating and hazardous gases only in ventilated rooms Hazard communication
Excess or empty cylinders present Respiratory protection
Chemicals Hearing conservation
Flammables stored in flammable storage cabinet Formaldehyde monitoring
Peroxide formers double-dated (received and opened) Ethylene oxide monitoring
Chemicals properly segregated Anesthetic gas monitoring
Hazardous chemicals stored above eye level General engineering controls
Chemicals stored on the floor Laboratory airflow is negative to general occupancy, corridor and office
Chemical containers left open areas
38 All solutions properly labelled Cup sinks or drains acting as vents 39
Mercury thermometers in use Sink available for hand-washing
Refrigerators/freezers/cold rooms Exposed machine parts (pulleys, gears)
Food for human consumption present Vacuum line has filters and traps on laboratory benches
Flammables in explosion-proof/-safe units labelled externally if Backflow hazards to water supply
containing carcinogens, radioactivity and/or biohazards Distilled water systems in good condition
Cold-room has emergency release Active and effective arthropod and rodent control program
Electrical equipment General practices and procedures
Extension cords present Food for human consumption stored outside the laboratory
Outlets earthed/grounded and with proper polarity Microwave oven(s) clearly labelled “No Food Preparation, Laboratory
Connections by sinks, under showers, etc. Use Only”
Equipment with frayed or damaged wiring Eating, drinking, smoking and/or applying of cosmetics occurring in the
laboratory
Overloaded outlets or electrical strips
Pressurized glass containers taped or shielded (i.e. vacuum traps)
Power strips mounted off the floor
General practices and procedures
Proper fuses in conduits
Mouth pipetting prohibited
Electrical outlets near water sources meet local codes
Mechanical pipetting devices available and used
Earths/grounds present on electrical cords
Protective laboratory clothing stored separately from street clothing
Portable space heaters
General laboratory housekeeping
Personal protective equipment
Glass containers stored on the floor
Eyewash available in laboratory
Trip hazards evident
Safety shower available
Clean absorbent pads on work surfaces
Personal protective equipment available (gloves, gowns, goggles, etc.)
Broken glassware handled by mechanical means (brush and dustpan,
Occupants properly attired
tongs, etc.)
Laboratory coats, gowns, smocks, gloves and other personal protective
Fire protection
clothing not worn outside the laboratory
Fire suppressor gas system heads free and unobstructed
Personal protective equipment available for cryogenic storage
Open penetrations in walls, ceiling, floor, etc.
Waste management
Wiring or tubing through door openings
Evidence of improper waste disposal
Minimum passage width of 1 m in laboratory
Wastes segregated in proper containers
Storage observed on ductwork or light fixtures
Chemical waste containers tagged, labelled, dated and kept closed
Heated constant temperature baths
Chemical waste containers appropriately handled and stored
Equipped with low water level and overheat shutoff
Sharps containers used and disposed of properly
Constructed of non-combustible materials
No trash on floor
Waste disposal procedures posted in laboratory
Occupational health and safety programs available
 Biological safety cabinet (BSC) Location: : Gloves worn when handling infectious material or contaminated
Brand : equipment
Type : Face protection provided when working outside the BSC with infectious
material
Serial
no.: Hands washed after removing gloves, after working with infectious
agents, before leaving the laboratory
Yes No N/A
Antimicrobial agent available for immediate first aid
BSCs located away from doors, heavily travelled areas, etc. to maintain
air flow Practices
Certification within last year BSC used when potential for creating infectious aerosols/splashes exists
BSC surface wiped down with appropriate disinfectant at beginning Biosafety manual prepared and adopted
40 41
and end of each procedure Personnel read, review and follow the instructions on practices and
Front grill and exhaust filter unobstructed procedures, including safety or operations manual (required for all
personnel annually)
Open flames used inside cabinet
Procedures performed so as to minimize aerosols/splashes
Vacuum lines have in-line filters and disinfectant traps in use
Needle-locking syringes/single-use needle syringe units used with
BSC compromised by room air or location
infectious agents
BSC used when there is potential for creating aerosols
Centrifuge cups and rotors opened only in a BSC
Laboratory
Infectious specimens transported outside a BSC in approved containers
Access limited and restricted to authorized personnel following approved transport regulations
Entry limited to personnel advised of all potential hazards Facility
Biohazard sign posted on laboratory door as appropriate Hand-washing sink available near laboratory exit
Information on sign accurate and current Laboratory separated from unrestricted traffic flow in building
Sign legible and not defaced Access to laboratory through an anteroom with self-closing doors
All doors closed All penetrations in laboratory sealed or sealable for decontamination
Decontamination Room exhaust air single-pass and exhausted away from occupied areas
Decontaminant specific to the organism(s) in use Controlled ventilation system to monitor directional airflow available
All spills and accidents involving infectious materials reported to the Personal protection
laboratory supervisor
Closed-front gowns worn in laboratory
Appropriate decontaminant used during spill clean-ups
Protective laboratory clothing worn only in laboratory areas
Work surfaces decontaminated before and after each procedure, daily
Hand-washing sink foot, elbow or automatically controlled
and after spills
Hand protection
Handling of contaminated waste
Double gloves worn when handling infectious material, potentially
Infectious waste containers properly used
contaminated equipment and work surfaces
Containers not overfilled
Respiratory protection
Containers properly labelled and closed
Respiratory protection worn by all personnel in the laboratory when
Culture stocks and other regulated waste properly decontaminated aerosols are not safely contained in a BSC
before disposal
Practices
Materials decontaminated outside the laboratory transported in closed,
Mucous membrane protection provided when working with infectious
durable, leakproof containers according to local rules and regulations
material outside a BSC
Mixed waste biologically decontaminated prior to disposal as chemical
Personnel advised of special hazards associated with the agent(s)
or radiological waste
Personnel required to read and follow all instructions on practices and
Personal protection
procedures, including safety or operations manual
Laboratory personnel reminded of appropriate immunizations/tests for
Personnel received annual updates/additional training for procedural
agents handled
changes
Appropriate medical services contacted for medical evaluations,
All contaminated waste autoclaved prior to disposal
surveillance and treatment of occupational exposures
 Annexure-7: BSL-3 Construction related information Annexure-8: Conceptual BSL-3 drawings
CIVIL WORK Humidistat [Diagrammatic depiction, how the lab area can be accommodated if the proposed lab is in an already
Doors Pan type humidifier existing building]
Windows CONTROLS
Walls & Ceiling Panels Air Handling Unit Controls
Epoxy Based Joint Less Flooring & Covings Condensing Unit Controls
Plumbing & Sanitary Installations Centrifugal Blowers
ELECTRICAL & ASSOCIATED WORKS Blower Drive Assembly
42 Internal Electrification of Building 43
MOTOR & SWITCHGEARS & MOTOR STARTERS
Main/Sub Distribution MV Panels Control Panel
Telephone System Cable Termination
Cables Indication
Earthing Subsidiary Panels
Fire Detection & Alarm System Contactor Starters
HVAC & ASSOCIATED WORKS Squirrel Cage Induction Motors
System Design DUCT WORK & OUTLETS
Drawings Duct materials & Installations
HVAC System based on of design Dampers
Operating Rooms / Zone Pressure Grilles & Diffusers
WATER CHILLING MACHINE Testing
Water Chilling Unit PIPE WORK
Compressor Pipes
Condenser (Air Cooled Type) Fittings
Chiller Flanges
Refrigerant Piping Valves
Motors & Starters Balancing Valves
Refrigerant Circuit Accessories Strainers
Control Panel Jointing
AIR HANDLING UNITS Hangers & Supports
WATER CIRCULATION EQUIPMENT Sleeves
Monobloc Type Pump Set Insulation Materials
Pump Accessories Chilled/Hot Water Piping
FILTERS Walls & Ceiling Acoustic Treatments of Plant Rooms & A.H.U. Room
Prefilters, Fine Filters & HEPA ELECTRIC WIRING
Filter Plenums Control Wiring
HEATING & REHEATING SYSTEM Compressors Condensers/Chillers/Evaporators/Pumps etc.
Electric Heaters & Heater Frames Air Handling Units
Contactors Piping System
Heating Thermostats Duct Work
 44
Facility Type (Areas of Primary Eye- Hand Shower On-site Ventilation Ventilation Air Effluent
Utilities) Containment wash wash Autoclave changes Decontamination
100% Fresh Recirculatory /Hr. System
air (30% Fresh [ACPH]** Chemical Steam
air)
Type Type
BSL-2, Type-1 (Basic laboratory Class II A2 * * NM NM [vertical As per risk May be 4-6 NM NA
for laboratory diagnosis autoclave assessment considered
[serological & molecular] for may be as per risk
RG-2 agents) considered] assessment
BSL-2, Type-2 (Laboratory with Class II A2 * * NM NM [vertical As per risk May be 4-6 NM NA
facility for propagation of autoclave assessment considered
infectious agents [serological may be as per risk
& molecular] for RG-2 agents) considered] assessment
BSL-2, Type-3 (Laboratory Class II A2 * M NM NM [vertical As per risk May be 4-6 M* NA
with facility for propagation autoclave assessment considered
of infectious agents in vitro may be as per risk
& in vivo for RG-2 agents) considered] assessment
+ equipped for providing
diagnosis for MDR & X-TB
ABSL-2, (Laboratory with Class II A2 * M NM NM [vertical As per risk May be 4-6 M* NA
facility for maintaining autoclave assessment considered
arthropods colony and may be as per risk
performing vector biology considered] assessment
work on infectious agents
of RG-2 agents) + equipped
for processing pools for the
surveillance of routine public
health vector borne diseases
like Dengue, Chikungunya,
Scrub typhus etc.
Table-1: Categories of laboratory and secondary barrier requirements

BSL-3, Type-1 (Basic laboratory Class II A2 M M NM NM [vertical M* NA 8-12 M* NA

for laboratory diagnosis autoclave
[serological & molecular] for may be
RG-3 known public health considered]
disease agents)
BSL-3, Type-2 (Laboratory Class II A2 M M NM* NM [vertical M NA 8-12 M* NA
with facility for propagation autoclave
of infectious agents in vitro may be
& in vivo for RG-2 agents) considered]
+ equipped for providing
diagnosis for MDR & X-TB
BSL-3, Type-3 (Laboratory Class II A2 M M NM* NM [vertical M NA 8-12 M*
for laboratory diagnosis and autoclave
with mandate of in vitro may be
propagation of specific considered]
infectious agents for RG-2
related to DNA recombination
technologies or RG-3 / MDR &
X-TB agents)
BSL-3, Type-4 (Laboratory for Class II A2 + M M M NM [vertical M NA 8-12 NA M
laboratory diagnosis and with IVC autoclave
mandate of in vitro and in may be
vivo propagation of specific considered]
infectious agents for RG-3
related to DNA recombination
technologies or RG-3 /MDR
& X-TB agents). Animal with
experimentation with RG-3
agents up to mice, chickens
and rodents.
45
 * Preferred, however depends on risk assessment for mandated work

Chemical Steam
Decontamination

Type
NA: Not applicable

M
NM: Not mandatory, depends on risk assessment for mandated work
Effluent

System

M: Mandatory
Type
NA

NA
**: Air Changes depends on risk assessment for mandated work
[ACPH]**
changes

NB: If mandate of the laboratory includes processing & propagation [in vitro] of unknown samples
10-12

10-12
from known or unknown high-risk group agents’ outbreak/ unusual outbreaks or regional VRDL: it is
/Hr.
Air

advised to follow BSL-3, Type-5 standards. However: If mandate of the laboratory includes processing &
46 propagation [in vitro & in vivo] of unknown samples from known or unknown high-risk group agents’ 47
100% Fresh Recirculatory

outbreak/ unusual outbreaks or regional VRDL: it is advised to follow ABCL-3 standards.

(30% Fresh
Ventilation Ventilation

Table 2. Onsite equipment in the laboratory

air)

M Equipment Remarks
Biosafety Cabinet Class-II (A1/A2/B1/B2) or Class-III, based on risk assessment
Autoclave Yes
Size and configuration shall be based on the program/laboratory’s
air

needs. Sterilization/ decontamination cycles and options are program

driven. Autoclave integrated with the containment barrier of the BSL-
NM [vertical

NM [vertical
considered]

considered]
Autoclave

autoclave

autoclave

3 laboratory shall provide bio-seal.

Shower On-site

may be

may be

Dunk Tanks & Pass Boxes Size and location of dunk tank/s and pass boxes shall be determined
by the program and shall be integral to the containment barrier. The
depth of partition must exceed the expected maximum pressure
differential.
Chillers for HVAC Capacity will depend on the total area of the facility.
M

*Boiler facility may be required if autoclaves operate on steam

Hand
wash

M
Containment wash
Eye-

M
Class II A2 +

Class II A2 +
Primary

IVC

IVC
related to DNA recombination
laboratory diagnosis and with

ABSL-3, (Laboratory for vector

biology experimentation and
for the surveillance of agents

experimentation facility with

BSL-3, Type-5 (Laboratory for

agents for RG-2 level related

vivo propagation of specific

technologies with facility to

& X-TB agents). Animal with

with RG-3 agents equipped

technologies or RG-3 /MDR

experimentation with RG-3

of vector borne diseases of

arthropods with infectious

maintain small animals for

mandate of in vitro and in

infectious agents for RG-3

RG-3 level or dealing with

experimentation facility
Facility Type (Areas of

to DNA recombination
with animal challenge

RG-3 agents)
Utilities)

agents
 Table-3: Thumb-Rule Calculations for Understanding
Estimated Budget of Proposed Laboratory Includes: Other materials, 300 f2 500 2,70,000
Item Work description Qty Unit Unit rate** Amount Chilled/Hot Water Piping,
Doors 12 Nos. 32,000 3,84,000 Walls & Ceiling Acoustic
Insulation
Treatments of Plant Rooms
Windows 5 Nos. 15,000 75,000
& A.H.U. Room, Acoustic
Walls & Ceiling Panels 2400 f2 3,000 72,00,000 Treatment Installation etc.
Civil Work Epoxy based joint less 243 m2 900 2,18,700 ++: Cost will depend on 1 Job 25,00,000 25,00,000
flooring & covings Building the area of the laboratory,
Plumbing & Sanitary 1 Job 2,20,000 Management which company? & critical
Installations System requirements of the specific
Internal electrification of 2,50,000 laboratory.
48 building Specialized ++: Depends on the 49

Main/Sub Distribution MV 3 Nos. 1,500 4,500 Equipments & requirements of the

Panels Systems laboratory.
Electrical &
Associated Telephone System 6 Nos. 2,000 12,000 Hot Water ++: Cost will depend on the 1 Job 3,50,000 3,50,000
Works Cables Shower number of shows required in
System the laboratory, which make?
Earthing 2 Nos. 18,000 36,000
++: Cost will depend on the 1 No. 4,00,000 4,00,000
Fire Detection & Alarm 15,00,000 15,00,000 Hot Water capacity required depending
System Generator on the number of showers in
System Design the laboratory, which make?
Drawings ++: Cost will depend on the 4 Nos. 35,000 1,40,000
HVAC System based on Bio-Safety number of biosafety doors
HVAC & the design and area of the Doors required in the laboratory,
Associated laboratory which make?
Works Operating Rooms / Zone ++: Cost will depend on 1 No. 40,000 40,000
Pressure Double Door the number of Double Door
++: Based on the design and Pass Box Pass Box required in the
area of the laboratory. laboratory, which make?
Water Chilling Unit [based on 50 Tr 40,000 20,00,000 Double Door ++: Cost will depend on the 1 No. 60,000 60,000
the design and area of the Pass Box number required & which
laboratory] (Dynamic make?
Compressor Type)
Condenser (Air Cooled Type) Steam ++: Depends on the 1 No. 12,00,000 12,00,000
Water Chilling Autoclave / requirements of the
Chiller
Machine Sterilizer laboratory.
Chiller Piping 15,00,000
++: Depends on the 3 No. 7,50,000 2,25,000
Motors & Starters 3 Nos. 30,000 90,000 Biosafety
requirements of the
Cabinet
Refrigerant Circuit laboratory.
Accessories Pre-Fabricated ++: Depends on the 574 f2 3,000 17,22,600
Control Panel 25,000 25,000 Shower requirements of the
Air Handling Air Handling Units 18 Tr 35,000 6,30,000 Cubicle laboratory.
Units CCTV ++: Depends on the area 1 Job 10,00,000 10,00,000
Water MoNobloc Type Pump Set 2 Nos. 20,000 40,000 System for & the requirements of the
Circulation Surveillance laboratory.
Pump Accessories
Equipment Total 82,05,900 2,29,41,800
Prefilters 3 Nos. 3,000 9,000
Fine Filters 3 Nos. 4,000 12,000
**=Unit cost for each item is non-realistic. These are given only to understand the process of
Filters High Efficiency Particulate Air 6 Nos. 38,000 2,28,000 calculation and represent only rough estimates. However, many items are not available on government
(HEPA) filters e-procurement sites like CPWD Manual Site etc. These can also be obtained from local contractors or
Filter Plenums 6 Nos. 1,00,000 6,00,000 may be searched on the websites.
50 51
 INDIAN COUNCIL OF MEDICAL RESEARCH

Department of Health Research

Ministry of Health & Family Welfare
V Ramalingaswami Bhawan, Post Box 4911,
Ansari Nagar-AIIMS, New Delhi – 110029 Delhi, India
Website: [Link]

ICMR 2019