Campylobacter ELISA Kit

Catalog No. ABIN997077

Product Details

Target: Campylobacter

Reactivity: Campylobacter

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Purpose: Campylobacter assay is an enzyme-linked immunosorbent assay (ELISA) that may be used to screen food products for the presence of thermophilic Campylobacter antigen.

Analytical Method: Qualitative

Application Details

Comment:

Quality Control:
The Positive and Negative Controls must be run each time the assay is performed. For a valid run, the Negative Control must be below 0.10 ODs and the Positive Control greater than 0.5 OD units. If either Control is out of range, do not use the kit and contact service.
Problem: Negative control has substantial color development. Correction: Washings were insufficient. Repeat test with more vigorous washings.

Plate: Pre-coated

Reagent Preparation:

Wash Buffer - Remove cap and add contents of one bottle of Wash Concentrate to a squeeze bottle containing 475 mL of DI water. Swirl to mix. Squeeze bottle should have a narrow tip to optimize washings.
MEDIUM PREPARATION
Bolton's Media: Acumedia catalog #7526 or equivalent - Follow manufacturer's instructions.
SAMPLE PREPARATION
Follow procedures listed in the Food and Drug Administration Bacteriological Analytical Manual (BAM- reference #5). Boil samples for 10 minutes and allow to cool before testing.

Assay Procedure:

1. Break off the required number of wells (number of samples plus 2) and place in strip holder.
   2. Add 100 μL of the negative control to well #1 and 100 μL of the positive control to well #2.
   3. Add 100 μL of the test sample to the appropriate well.
   4. Incubate at room temperature (15 - 20 °C) for 30 minutes, then wash.
   5. Add 2 drops of Enzyme Conjugate (red solution) to each well.
   6. Incubate for 15 minutes, then wash. Slap out excess fluid against an absorbent towel.
   7. Add 2 drops of Chromogen to each well.
   8. Incubate for 5 minutes.
   9. Add 2 drops of stop solution to each well. Mix wells by gently tapping the side of the strip holder with index finger.
   10. Read results visually or at 450/620-650 nm.
   Each washing consists of dumping the contents of the wells into an appropriate container with disinfecting solution (e.g. 3 % bleach in water) and using the diluted wash buffer to fill in each well, shaking out the contents and refilling the wells for a total of 3 times. Samples with sticky particulate matter may require more thorough washing than other samples. The potential exists for false positive results if the sample is not thoroughly washed from the well before addition of subsequent reagents. Only one set of controls is required per run. Read results within 4 hours from addition of Stop Solution. All incubations are at room temperature (15-25 °C).

Calculation of Results:

VISUAL Positive: Any sample well that has significant and obvious yellow color. Negative: Any sample well that does not have significant and obvious yellow color.
NOTE: The negative control, as well as some samples, may show some slight color. A sample well must be obviously darker than the negative control well to be called a positive result.

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment: Store at 2-8 °C

Target Details

Target: Campylobacter

Background: Infection by thermophilic Campylobacter species is a leading cause of human gastroenteritis. Of the various species of Campylobacter, C. jejuni, C. coli and C. lari, are the species most often associated with human illness. Campylobacter are often passed to humans through the handling or consumption of contaminated food, particularly foods of animal origin. Recently, human infection with Campylobacter has been implicated in the induction of Guillain-Barré Syndrome (GBS) and reactive arthritis. GBS is a debilitating and potentially fatal neurological disease that produces paralysis. Campylobacter species are gram negative, motile curved or spiral rods that require highly specialized growth conditions. Typical cultivation entails pre-enrichment and enrichment steps in broth, followed by isolation on a selective solid medium. Of particular importance in the cultivation of Campylobacter is the requirement for a microaerobic atmosphere. Cultivation of thermophilic Campylobacter species from foods involves multiple media steps, and thus many days. The Diagnostic Automation, INC. Campylobacter Microwell ELISA allows rapid testing (less than an hour) of the enrichment broth, thus eliminating the need for the plating and identification steps used in the standard methods.