MYCOLOGY AND [TRANS] CHAPTER 1:

VIROLOGY

GENERAL FEATURES OF VIRUSES  cell lysis- naked

- Obligate intracellular parasites unable to self-replicate. Once  budding- enveloped
inside living cells, viruses induced the host cell or synthesize virus EBV CD3 receptor
particles HIV CD4
- Genome is either DNA or RNA Rabies Acetylcholinesterase
- Haploid (only one copy of their genome except; retrovirus (Diploid) SARS-COV ACE 2
Infectious Influenza Sialic Acid
- Do not have a system to produced ATP
- Viruses ranges in size from 25 to 270nm
- Maybe Virulent (cause death e.g. Rabies) or Temperate (doesn’t VIRAL REPLICATION
caused death) - Formation of biological viruses during the infectious process in target
host cell
- Viruses must get into the cell before cell replicate occurs
- DNA viruses assembled in the NUCLEUS and RNA viruses
developed solely in CYTOPLASM
- Greatly varied and depend on the type o genes involved in them

i. Attachement
VIRAL STRUCTURE - Attachment of virus to host cell receptor
Virion - Complete virus particles - Must have a strong interaction between virus and host cells
- infectious unit - Complementary parts of the virus is called LIGAND while the host
Nucleic Acid - DNA or RNA (genome) cell surface is called RECEPTOR
- Single or Double
- Linear or Nuclear ii. Penetration
Capsid - Protein coat that encloses genetic material -Virus enters host cell by direct penetration, endocytosis or fusion
- May be helical (rod like) Icosahedral with cell membrane
(cuboid) or complex (i.e. Poxvirus) -Direct penetration- Naked
- Composed of protein subunits called -Endocytosis or Fusion- Enveloped
capsomeres
- Protects nucleic acid, enables virus to attach Mode by which viruses penetrate:
to and enter host cell
a. Naked virus direct penetration
Nucleocapsid - Nucleic acid + capsid
b. Virus with enveloped penetrates by endocytosis
Enveloped - Outer membrane surrounding capsid in some c. Virus with enveloped penetrate by fusion of cell in membrane
viruses; Aids in attachment to host cell
o ENVELOPED VIRUS: Ether Sensitive
iii. Uncoating
o NAKED VIRUS: Ether Resistant
iv. Macromolecules
v. Assembly
-Attachment of virus to host cell receptor vi. Maturation
Attachment/ -Virus exhibit tropism (ability of virus to vii. Release
Adsorption recognize and attach to limited number of
cells)
-Virus enters host cells by direct penetration,
endocytosis or fusion with cell membrane
Penetration  direct penetration- naked
 endocytosis or fusion cell-enveloped
Uncoating -Loss of capsid
-Genome enters cytoplasm (for most RNA
viruses) or nucleus (for most DNA viruses)
Macromolecular -Production of nucleic acid and protein polymerase
Synthesis -transcription and translation occurs
Assembly -Structural proteins, genome, viral enzymes are
assembled into virus particles
-Envelops required during viral ‘budding’ from
hose cell membrane
-Occurs after cell lysis or by virus particle budding
Release from cytoplasmic budding

MONTECALVO, SHARMAINE A. BSMT4

 MYCOLOGY AND [TRANS] CHAPTER 1:
VIROLOGY
Rubella german measles
HSV,VSV Tzank smear
Poxvirus largest DNA virus

CRITERIA OF VIRUS CLASSIFICATION: SPECIMEN COLLECTION AND TRANSPORT

1. Nucleic acid composition
a. DNA or RNA - Timing of collection- during acute phase
b. SINGLE or DOUBLE STRANDS - Collect in sterile, leaked proof, non-breakable container
2. Capsid morphology - Swabs: Dacron, Rayon or other polyester tips with plastic
3. Presence or absence of enveloped or aluminium shafts
4. Site of virion assembly o Calcium alginate, cotton, wood- inhibitory for some
viruses
TAXONOMY: o Swab specimen should be emulsified in viral transport
 Virus family- “idae’ medium before transport to the laboratory, especially if
 Subfamilies- “inae” transport will occur at room temperature and requires
 Genera- Virus longer than 1 hour
- Specimen should not be allowed to sit at room temperature or higher
SPECIMEN COLLECTION AND TRANSPORT temperatures
Infection site Specimens Common viruses - Specimen should be kept cool (4C) and immediately transported to
CSF, Enteroviruses (#1 common the laboratory
CNS Throat swab, cause of aseptic meningitis) o If delayed, specimen should be refrigerated
Brain tissue, Blood HSV, - Process specimen within 12 to 24 hours of collection
Arboviruses o If not possible, store specimen at 4C (up to5 days)
Eye Conjunctival, HSV,
o Keep at 20C, preferably -70C (6 days or longer)
Corneal scrapping Adenovirus
o Specimen for freezing should first be diluted or emulsified in
Genital swab, HSV,
Genital tract Vesicle swab or fluid, HPV (#1 common cause of viral transport medium
Lesion biopsy cervical cancer)
adults: Noroviruses, INTENSITY IN FITC FLUORESCENCE
Adenoviruses, Enteroviruses Faint yet unequivocal apple green fluorescence 1+
Stool, Apply green fluorescence 2+
GI tract Rectal swab children: Rotavirus (#1 Bright apple green fluorescence 3+
common cause of diarrhea), Brilliant apple green fluorescence 4+
Adenovirus
Nasal aspirate, Influenza A and B,
DETECTION METHODS
Respiratory Throat swab, Parainluenza virus,
Tract Nasopharyngeal swab, Adenovirus,
BAL, RSV, HMPV, I. MICROSCOPY
Lung biopsy Rhinovirus, Enteroviruses
HSV and VSV (Tzank  Light Microscopy
smear), Measles, - for identification of large virus (i.e. Poxvirus)
Skin Vesicle blood Rubella (German measle), - TZANK SMEAR: HSV, VSV
Enteroviruses, Parvovirus - KOILOCYTES- HPV
B19
 Attypical aquamous cells with large nucleus surrounded by non-
Urinary Adenovirus,
staining halo
Tract Urine HSV,
CMV
Blood Blood CMV  Electron microscopy
- rarely used, labor intensive, expensive, relatively insensitive
SIDE NOTES - to assess viral morphology
Rhinovirus Can get through rain (kapag nabasa ka sa ulan hindi - most useful for detecting gastroenteritis virus that cannot be
balance ang temperature sa katawan detected by other methods (e.g. astroviruses) and encephalitis causing
Enterovirus common cause of aseptic meningitis viruses that are undetectable with cell culture (HSV, Measles,Jc)
HPV common cause of cervical cancer - Stains for viruses of Electron Microscopy:
Rotaviruse common cause of diarrhea in inants/children  Potassium Phosphotungstate

MONTECALVO, SHARMAINE A. BSMT4

 MYCOLOGY AND [TRANS] CHAPTER 1:
VIROLOGY
 Uranyl Acetate

Cytomegalovirus Owl’s eye

Herpes Simplex Virus Cow dry Type A
Poxvirus Guarnieri Bodie (small pox)
Poxvirus Mollusum Bodies (Mollusum contangiosum)
Rabies Negri Bodies
Yellow fever Torres Councilman
 Phase-contrast microscopy
- To see viral inclusions found within infected

II. SEROLOGY
- More sensitive
- Detects antibodies in serum
- Evaluating immune status
- Diagnosing viral infections

 Classical Technique: Hemagglutination and Compliment fixation

 Older technique: RIA (toxic)
 Newer Technique: Western Blot and EIA

SNOW DROP:
Southern Blot DNA
Northern Blot RNA
O O
Western Blot PROTEIN

III. MOLECULAR
- Faster and more sensitive
- PCR, Real-time PCR, Branched DNA, Nucleic Acid Hybridization
- Detects virus that cannot be cultured

IV. CULTURE
- Deferent viruses grow in different cell lines
- No single cell types grows all viruses
- Several typed should be inoculated
- Incubate inoculated cell cultures immediately at 35C
- Growth may take 1 to 28 days

GRADING OF CPE
Intensity Interpretation
- Uninfected monolayer
1+ 1 to 25 of monolayer exhibits Cytopathogenic
effect
2+ 26 to 50 of monolayer exhibits Cytopathogenic
effect
3+ 51 to 75 of monolayer exhibits Cytopathogenic
effect
4+ 76 to 100 of monolayer exhibits Cytopathogenic
effect

MONTECALVO, SHARMAINE A. BSMT4