VIP® for Listeria

AOAC Official Method 997.03

AOAC Performance Tested Method 060801

GENERAL DESCRIPTION (b) Inactivate microorganisms at 100°C (95-105°C) for 5 min

VIP® for Listeria is a single-step visual immunoassay for the in a water bath, autoclave or dry heater.
detection of Listeria in food and environmental samples. Each (c) Cool tubes to 25-37°C before testing. Tubes that have
device contains a proprietary reagent system, which forms a been inactivated can be stored for up to 4 days at 2-8°C
visually apparent antigen-antibody-chromogen complex if Listeria prior to testing.
is present. The test is intended for use by laboratory personnel
with appropriate microbiology training. TEST PROCEDURE

Part No’s: 600120 (20 tests), 600150 (50 tests) (a) Open the sealed pouch containing VIP units and remove
the required number of tests. One device is necessary
KIT COMPONENTS for each test portion. VIP units may not be reused.
Each VIP for Listeria kit contains the following: Reseal unused VIP units in pouch containing desiccant.
VIP for Listeria test devices Store at room temperature (15-25°C)

EQUIPMENT / MATERIALS REQUIRED (b) Enriched broths should be equilibrated to 25-37°C prior
to running the test.
Other necessary materials not provided include:
Media per Appendix A (c) Vortex mix contents. Transfer 0.1 mL of inactivated
Autoclave BLEB or DFB to sample addition well. Avoid transferring
Vortex mixer particulate matter to the device.
Analytical balance, tolerance ± 0.2 g (d) Incubate device at room temperature (15-25°C) for 10
Stomacher / Masticator machine min.
Stomacher-type bags with filter or equivalent
Incubator capable of maintaining 29-31°C RESULTS
Water bath capable of maintaining 95-105°C or Note: Examine test unit at 10 min. Beyond this time faint
equivalent (e.g. autoclave with flowing steam, dry heater) lines may develop because of non-specific color development
Micropipette(s) capable of delivering 0.1 mL and 1.0 mL and should be disregarded.
SAMPLE PREPARATION (a) Examine VIP unit for the presence of distinct detection
A. Test Portion Preparation & Enrichment lines in both test sample and test verification zone. Lines
should be dark when contrasted with white background
Two Step Enrichment Protocol (AOAC OMA 996.14)- and should extend across the zone. Intensity of test
for meat, poultry, seafood, vegetables, dairy products, eggs, sample and test verification lines may differ. Absence of
pasta, animal meal, nuts and environmental samples. a test verification line indicates an invalid test result.
(a) Food samples Add 25 g test portion to 225 mL of Contact BioControl Technical Services at (800) 245-
modified Fraser Broth with lithium chloride (mFB+LiCl) 0113.
(Appendix A). Stomach / masticate for 2 minutes and (b) Test sample is considered positive when lines are present
incubate 28 h in the test sample zone and in the test verification zone.
(26-30 h) at 30°C (29-31°C).
(c) Test sample is considered negative when a line is present
Environmental samples Add 60 mL of mFB+LiCl to in the test verification zone and no line is seen in the test
sample bag containing environmental sponge sample. sample zone.
Ensure that the sponge is oriented horizontally in the
sample bag. If using a swab, add environmental swab (d) Positive and negative control cultures should be run to
sample to 10 mL of mFB+LiCl. Mix well and incubate 28 h familiarize the analyst with the results interpretation.
(26-30 h) at 30°C CONFIRMATION
(29-31°C).
Presumptive positive samples should be confirmed from the
(b) Transfer 1 mL incubated mFB+LiCl to 9 mL Buffered retained BLEB or DFB tubes via either:
Listeria Enrichment Broth (BLEB) (Appendix A). Mix
thoroughly with vortex mixer. Incubate 24 h (22-26 h) at US FDA. 2003. Bacteriological Analytical Manual online, chapter
30°C (29-31°C). 10. [Link]

Single Step Enrichment Protocol (AOAC PTM [Link]) USDA-FSIS. 2008. Microbiology Laboratory Guidebook, chapter
- for meat, poultry, seafood, vegetables, dairy products, and 8.[Link]
environmental samples. k/[Link]

(a) Food samples Add 25 g test portion to 225 mL of Demi- STORAGE CONDITIONS
Fraser Broth (DFB) (Appendix A). Stomach / masticate for Store VIP for Listeria kit components at room temperature
2 minutes and incubate 48 h (46-54 h) at 30°C (29-31°C). (15-25°C). Do not refrigerate.
(b) Environmental samples Add 100 mL of DFB to sample Reseal unused VIP units in pouch with desiccant pack. The
bag containing environmental sponge sample. Ensure that moisture indicator line on the desiccant pack must be blue.
the sponge is oriented horizontally in the sample bag. If
using a swab, add environmental swab sample to 10 mL of Kit expiration is provided on the product pouch label.
DFB. Mix well and incubate 48 h (46-54 h) at 30°C (29- PRECAUTIONS
31°C).
Assurance EIA for Listeria must be used as described herein.
Note: Retain original BLEB or DFB tubes under This product is not intended for human or veterinary use.
refrigeration (2-8°C). Use for confirmation of presumptive Do not use test kit beyond expiration date indicated on the
positive results (section E). label. Do no mix reagents from different Assurance kit lots.
B. Sample Inactivation Avoid contact with eyes, skin, and clothing. Avoid swallowing
(a) Vortex mix incubated sample. Transfer 1.0 mL incubated or taking product internally.
BLEB or DFB to a clean test tube.
Decontaminate materials by autoclave, bleach, etc., in
accordance with good laboratory practices and in accordance
with local, state and federal regulations. Waste may be
contaminated with Listeria which is potentially hazardous to
human health. All biohazard waste should be disposed of
appropriately.
Pregnant women, elderly, and potentially
immunocompromised individuals must be prohibited from
laboratory rooms or areas where L. monocytogenes
enrichment, isolation, and identification procedures are in
progress. Although a properly sanitized laboratory area
should not harbor Listeria, supervisors should use their own
discretion in allowing these high-risk individuals into these
areas.
PRODUCT WARRANTY
BioControl Systems, Inc. (BCS) warrants this product to be free from defects
in materials and workmanship, when stored under label conditions and used
as intended until the expiration date stated on the package. BCS agrees
during the applicable warranty period to replace all defective products after
return to BCS. BCS shall not have obligation under this Limited Warranty to
make replacements which result, in whole or in part, from negligence of the
Buyer, or from improper use of the products, or use of the product in a
manner for which it was not indicated. Buyer shall notify BCS of any
products which it believes to be defective during the Warranty period. At
BCS option, such products shall be returned to BCS, transportation and
insurance prepaid. BCS shall replace any such product found to be defective,
at no charge. Should BCS examination not disclose any defect covered by
the foregoing warranty, BCS shall so advise Buyers and dispose of the
product in accordance with Buyer’s instructions.

BioControl Systems, Inc. 2008 BioControl Systems, Inc., certifies that VIP for
®
Listeria meets all requirements specified in AOAC Official Method 997.03
Listeria monocytogenes and related Listeria spp. in Selected Foods, Visual
Immunoprecipitate Assay. BioControl and VIP are registered trademarks of
BioControl Systems, Inc. US patent #5,658,747, European patent
#0,759,172. Canada patent #2,190,109. Australia patent #680362. Japan
patent #2894841. Mexico Patent #230973. Other US and international
patents are pending. Printed in USA. All rights reserved.

APPENDIX A

Modified Fraser Broth with Lithium Chloride

(mFB+LiCl)
Suspend 55 g of Fraser Broth Base (FB) in 1 L deionized
water. Mix thoroughly until completely dissolved. Add 4 g
lithium chloride (LiCl) and stir until completely dissolved.
Autoclave at 121ºC for 15 min. Do not overheat. Do not
add ferric ammonium citrate additive to broth. Do not use
broth if precipitate forms. Alternately, prepare a 45% (w/v)
LiCl solution by dissolving 45 g LiCl in enough deionized
water to make a final volume of 100 mL. Filter sterilize
through a 0.2 µm filter. Add 2 mL sterile LiCl solution to 225
mL of sterilized FB. If using commercially prepared 8 M LiCl
solution, add 2.65 mL per 225 mL sterilized FB.

Buffered Listeria Enrichment Broth (BLEB)

Suspend 36.1 g Listeria Enrichment Broth in 1 L deionized
water. Add 8.5 g 3-3-(N-Morpholino) propanesulfonic acid
(MOPS) free acid and 13.7 g MOPS sodium salt. Mix
thoroughly. Autoclave at 121°C for 15 min.

Demi-Fraser Broth (DFB)

Suspend 55 g of Demi-Fraser Broth Base in 1 L deionized
water. Mix thoroughly until completely dissolved. Autoclave
at 121ºC for 15 min. Do not add ferric ammonium citrate
additive to broth.