HI

you can use selective isolation media Campylobacter Agar Base .

good luck

Isolation on Cam. Specific agar and Identification using Campylobacter API Kit ( Biomereux,Montalieu,France).

Also specific marker genes you can make PCR with specific primers but API Kits are more accurate.

Hi Ali,

What kind of test are you looking to do (biochemical or PCR) and how specific do you need to go? If you are just looking to identify your isolates as Campylobacter spp. there are a few simple biochemical tests you can perform. I recommend using the Oxoid O.B.I.S. Campy kit (Thermo Fisher, IDO800M). It performs the Gram lysis test and can also tell you if your suspect isolate can hydrolyse the substrate L-ananyl-7-amino-4-methylcoumarin using a simple colorimetric test (Hint: a negative reaction indicates the organism is a presumptive Campylobacter, Helicobacter or Arcobacter species).

Oxidase detection strips (Oxoid, MB0266A) are also cheap, simple to use and extremely useful for Campylobacter identification. It is used to detect the presence of the bacterial cytochrome oxidase enzyme and determines if the bacteria utilises oxygen with an electron transfer chain. Suspect Campylobacter colonies will turn a deep purple/violet colour after a few seconds. I recommend testing colonies that have been cultured on mCCDA or some other media that does not contain blood supplements though as the red blood cells contain cytochrome oxidase which can give rise to false positive results.

The above tests are relatively quick and, in the long term, are much cheaper to use than API biochemical kits. You could also subculture isolates onto two Columbia Blood agar plates and grow one plate aerobically at 41.5C and the other microaerobically at 25C. Campylobacter cells do not grow under these conditions.

If you want to be more specific and identify your isolates to the species level there is a PCR technique you can use that not only confirms your isolate is Campylobacter but also tells you if it is C. jejuni, C. coli, C. upsaliensis, C. lari or C. fetus subsp. fetus. You can read the paper and the methods here:

Article Colony Multiplex PCR Assay for Identification and Differenti...

The most common selective agar used is charcoal cefoperazone deoxycholate agar (CCDA). Plates are incubated at 37°C for 2 days in anaerobic jars or in a Campy gas atmosphere. The colonies of Campylobacter usually are gray, flat, irregular, and spreading in freshly prepared media.