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You are here:Home » Kits and Assays » Kits-ELISA (Food Safety), BioAssay™ » Campylobacter Antigen (In Food) BioAssay™ ELISA Kit

Campylobacter Antigen (In Food) BioAssay™ ELISA Kit

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Specifications

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.
Catalog #C1037-19
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 United States Biological 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. The United States Biological Campylobacter Microwell ELISA reduces time to result by several days.
Intended UseThe Campylobacter Antigen (In Food) ELISA Kit is an enzyme-linked immunosorbent assay that may be used to screen food products for the presence of thermophilic Campylobacter antigen.
Principle of the TestThe Campylobacter Antigen (In Food) ELISA is a double antibody (sandwich) assay that uses specific anti-Campylobacter antibodies coated to microwells. After addition of the sample and the enzyme conjugate, a positive reaction (indicating the presence of Campylobacter antigen) produces a deep blue color. Addition of the Stop Solution ends the assay and turns the blue color to yellow. The results may be read visually or with an ELISA reader.
Kit ComponentsC1037-19A: Microwell test strips, 1x96 wells and supplied with one holder. Coated with anti-Campylobacter polyclonal antibody.
C1037-19B: Enzyme Conjugate, 1x11ml. Peroxidase conjugated anti-Campylobacter polyclonal antibody with red dye and a preservative.
C1037-19C: Positive Control, 1x2ml. Campylobacter antigen in a buffered base.
C1037-19D: Negative Control, 1x2ml. Buffered base.
C1037-19E: Chromogen, 1x11ml. Tetramethylbenzidine (TMB).
C1037-19F: Wash Concentrate (20X), 2x25ml. Concentrated buffer and surfactanct with preservative.
C1037-19G: Stop Solution, 1x11ml. Phosphoric Acid.
Storage and StabilityStore reagents, strips and bottled components at 4°C. Unopened kit is stable for 6 months upon receipt.
Test Procedure1. Break off the required number of wells (number of samples plus 2) and place in strip holder.
2. Add 100ul of the negative control to well #1 and 100ul of the positive control to well #2.
3. Add 100ul of the test sample to the appropriate well.
4. Incubate at room temperature (15 to 25°C) for 30 minutes, then wash. 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.
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.
Interpretation of Results –VisualPositive: Any sample that has significant and obvious yellow color.
Negative: Any sample that does not have significant and obvious yellow color.
Interpretation of Results – OD ReadingsZero reader on air. Read all wells using a bichromatic reading with filters at 450nm and 620- 650nm.
Positive Add 0.10 OD unit to the value of the negative control. Sample OD values of 0.10 above the negative control indicates the sample contains Campylobacter antigen.
Negative Absorbance reading less than 0.10 plus the negative control indicates the sample does not contain detectable levels of Campylobacter antigen.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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