I am performing ELISA with the serum of dogs to detect IgG against Campylobacter. But all samples turn out positive and with very high ODs. That's impossible. Does anybody know a possible reason for that or a solution? What kind of cross reaktivity may exist. Or do the samples need to be treated before detetction. The dilution of 1:10 and 1:100 show the same results. Everything is positive and too high. The Standards and negative control are fine. Thanks for any advice
Dear Daniel,
your working conditions seem not optimal. In case you are developing an ELISA test, please try different conditions, regarding Ag con., primary and secondary Ab dilutions. If you are working with a commercial ELISA, the problem could be from washing; so you may need extra washing steps.
You can also try using 0.03 M urea in the serum diluent and incubate the plates at 37C for 1 hr then wash and add the secondary Ab. This may reduce the cross reactivity. Cross reactivity is often observe with canine sera.
Hope that is useful.
Elfadil
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