Reveal for Salmonella Test System

Neogen Corporation
Lansing, MI


INTENDED USE:

The Reveal for Salmonella test system permits rapid recovery of salmonella organisms in food and feed, allowing detection by the Reveal test device within 20 hours.



INTENDED USER:

Reveal for Salmonella test system is designed for use by personnel familiar with the appropriate techniques of working with salmonellas. Training is recommended for those without a basic knowledge of microbiology.



ASSAY PRINCIPLES:

This system utilizes REVlVE (TM) medium which presents salmonella organisms with readily available nutrients and other factors required for their survival and recovery from a stressed or injured condition. After which, selective enrichment promotes growth to detectable levels by the Reveal lest device which is described below.

A sample of enrichment culture is placed into the round sample port followed by enhancement reagent. Contents of the sample are wicked through the pad to a specimen reaction zone containing colloidal gold-labeled antibodies specific to commonly encountered salmonella serovars. Reactive salmonellas complex with the gold labeled antibodies and migrate through the support until they encounter a binding reagent zone which includes a second antibody specific to salmonellas. When this occurs a line forms in the test window due to the aggregation of the gold-labeled antibody complexes.

The rest of the sample continues to migrate until it encounters a second binding reagent zone which contains an antibody specific for the gold-labeled antibody. This results in the formation of a line in the control window. Regardless of whether or not the sample contains salmonellas, a line will form, ensuring the test is working properly.

REFRIGERATE POUCHES AT 2-8C; DO NOT FREEZE!


REAGENTS PROVIDED:

Test Kit Contents:

Reveal for Salmonella test device (20) pouched individually
Disposable transfer pipette (20)
Enhancement Reagent (1 dropper bottle each for and RV)



REVIVE Media Kit:

REVIVE media (20 bottles)
Stomacher-type bag (20)
Graduated Cup - sterile or sanitized



Selective Enrichment Media Kit:

Selenite Cystine (SC) or Rappaport Vaseiliadis (RV) Concentrates (20 bottles per package)
Stomacher-type bags
Graduated Cup - sterile or sanitized

See Table 1. Commodities List



MATERIALS REQUIRED BUT NOT PROVIDED

High Speed Blender (1 qt. jar), Stomacher machine or equivalent
Incubators or water baths (2): Capable of 37 and 43 + 1.0C
Balance
Timer
Distilled or Sterile water



PRECAUTIONS

1. Store Reveal test devices at 2-8C.
2. Allow Reveal test devices to warm to room temperature before opening pouch.
3. Store media at room temperature (18-30C)
4. Do not rehydrate REVIVE Media or selective enrichment media over six hours in advance.
5. Use of incubation times other than those specified may give erroneous results.
6. Sample bags must be closed loosely to allow air exchange during incubation which is vital to survival and antigenic expression by the Salmonella bacteria.
7. Distilled or sterile water should be brought to 43C before use. Deionized water must be boiled (sterilized) and cooled to 43C before using.
8. Test sample within 2 hours post incubation.



I. SAMPLE PREPARATION AND PRE-ENRICHMENT

A. STOMACHER OR HAND MIX METHOD: For dry or malleable foods (i.e. commodities such as liquids, dry goods, ground meats, feeds, grains, etc.)

1. Transfer contents of one (1) bottle of REVIVE to the Stomacher-type bag. Using the sterile graduated cup, add 200 mL of pre-warmed (43C) distilled or sterile water. Grasp bag tightly 2-3 inches from the top and mix vigorously until dissolved (about 1 minute).

2. Place 25 g of sample (sample must be room temp.) into the Stomacher-type bag containing the REVIVE medium. Grasp bag tightly at top and knead sample until homogenous. Shake bag vigorously using side-by-side motion to ensure complete mixing. Alternative: place bag in a Stomacher machine and "stomach" for two (2) minutes at normal speed.

Note: whole or pelleted dry commodities should be ground to a fine consistency before adding to REVIVE.

3. Close bag loosely (see caution #6) and place in suitable rack or support. Incubate at 37C for:
2 hours if using Selenite Cystine (SC) selective enrichment or
4 hours if using Rappaport Vassiliadis (RV) (See Table 1)



B. BLENDER METHOD - For sinewy or fleshy foods (i.e. commodities such as whole meats, seafood, salami, tissues, etc.)

1. Transfer contents of one (1) bottle of REVIVE to a sterile or sanitized dry blender jar. Using the sterile graduated cup, add 200 ml of prewarmed (43C) distilled or sterile water. Blend at high speed for 30 seconds to dissolve.

2. Place 25 g of sample (sample must be room temp.) into blender jar and cap tightly. Blend at high speed for one (1) minute or until sample appears well mixed.

3. Add contents of blender jar to the Stomacher-type bag and close loosely (see precaution #6).

4. Place bag in a rack or suitable container and incubate at 37C for:
2 hours if using Selenite Cystine (SC) selective enrichment
4 hours if using Rappaport Vassiliadis (RV) (See Table 1)



Table 1. Commodities List
(Grouped by recommended selective enrichment media)

Selenite Cystine (SC)

Rappaport Vaseiliadis (RV)

Beef Tissue Pork Tissue Beef Tissue Pork Tissue
Cake Mixes Poultry Rinse Crab Meat Powdered Milk
Chocolate Poultry Feed Fishmeal Processed Meats
Ground Beef Processed Meats Liquid Eggs Seafoods
Ground Chicken Soybean Meal Poultry Rinse Shrimp
Liquid Eggs Yeast    



II. SELECTIVE ENRICHMENT

A. RECONSTITUTION

1. Transfer contents of one bottle of the appropriate selective enrichment (Table1) to the bag provided. Using the sterile graduated cup, add 200 ml of prewarmed (43C) distilled or sterile water to the bag. Grasp bag tightly 2-3 inches from the top and mix vigorously until dissolved (about 1 minute).

Note: Keep reconstituted media at 43C until needed (not more than 6 hrs)

B. ENRICHMENT

1. Remove the REVIVE sample bag from the 37C incubator after the required incubation period (A3, B4) and place in a suitable rack or support. Add the 200 mL of 43C selective media to the sample bab. Shake bag vigorously using side-to-side motion to ensure complete mixing.

2. close bag loosely (see precaution #6) and place in suitable rack or support. Incubate at 43C for:
18 bours if using Selenite Cystine selective enrichment or
18hours if using Rappaport Vassiliadis enrichment



III. TESTING WITH REVEAL

A. Remove sample bag from the incubator and place in a suitable rack or support and allow contents to equilibrate to room temperature (18-30C). Test sample within two (2) hours post incubation. Allow device to warm to room temperature before opening pouch.

B. Remove Reveal test device from its foil pouch and place on a flat surface. Label each device with sample identification.

C. Using a transfer pipette provided, draw sample liquid up into stem portion and dispense three (3) free falling drops (about 100 ul) into the circular sample port of the device. As soon as the sample liquid is absorbed, add one drop of the appropriate Enhancement Reagent dropper bottle (SC or RV) into the sample port.

D. Observe test results within 15-20 minutes. Do not read after 20 minutes.



CONFIRMATION

If a sample Is positive, you may wish to confirm by plating the REVIVE selective medium sample on media recommended in the BAM or FSIS procedures (See Bibliography).

Example:
Using appropriate techniques, streak REVIVE selective media sample onto XLD, BS. HE and XLT4 agar plates and incubate 24 hours at 37C. Observe plates for typical Salmonella colonies. Colonies can then be used to inoculate TSI and LIA slants for further confirmation.

Note: Testing a new subsample using BAM or FSIS procedures may not yield the same result. Bacteria are not evenly distributed within a sample (especially in dry or solid samples) hence, the subsample may not contain the organism.



DISPOSAL:

Dispose Reveal test devices pipettes and media by one of the following methods:

1. Autoclave all components at I5 psi at 121C for one hour.

2. Place used test devices and pipette into the sample bag and add 50 mL liquid bleach (5.6% sodium hypochlorite). Allow to stand for a minimum of two (2) hours. Devices and pipettes can then be discarded and the liquid can be poured down the drain. Flush well with water.



CUSTOMER SERVICE:

Customer assistance and Technical Service for Reveal and REVIVE can be reached at the numbers shown below between 8 a.m. and 8 p.m. EST. Training on this product and all Neogen test kits is available.

Division

Phone Number

Feed, grain, nuts, and service labs 800-234-5333
Meat and Poultry Products 800-234-5333
Seafood Products 800-477-8201
International 517/472-9200



BIBLIOGRAPHY:

Bacteriological Analytical Manual (6AM), 8th Edition (1995), published and distributed by AOAC International, 481 N. Frederick Ave. Suite 500, Gaithersbung, MD 20877-2504 USA 301-924-7090



WARRANTY:

Neogen Corporation makes no warranty of any kind, either expressed or implied, except that the materials from which its products are made are of standard quality. If any materials are defective, Neogen will provide a replacement product. Buyer assumes all risk and liability resulting from the use of this product. There Is no warranty of merchantability of this product, or of the fitness of the product for any purpose. Neogen shall not be liable for any damages, including special or consequential damage, or expense arising directly or indirectly from the use of this product.

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