Sensitivity and specificity of a latex agglutination test for detection of calf enterotoxigenic Escherichia coli isolates in comparison with PCR
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Latex agglutination test is a very fast, accurate, and specific method that is used to diagnose antigens and antibodies which can be carried out in any place without need for any specific devices. In the present study, a single chain (scFv) recombinant monoclonal anti-K99 antibody was used in designing and producing a latex agglutination kit for the detection of enterotoxigenic Escherichia coli of calves colibacillosis. The purpose of this research was to primarily evaluate the sensitivity and specificity of the mentioned latex agglutination kit in the detection of the considered bacteria and to compare it with the results of PCR method. To carry out latex agglutination method, carboxyl latex particles were covered by the recombinant monoclonal antibodies of anti-K99 scFv. Then, these latex particles were mixed with the bacterial suspension purified from the diarrhea samples of the calves on the black cards of kit to observe agglutination. Sensitivity and specificity of the latex agglutination kit in the diagnosis of this colonization factor was evaluated and compared with the results of PCR test. Comparison of the PCR test results with those of the kit indicated that, under the laboratory conditions and on the purified bacteria, sensitivity and specificity levels of the latex agglutination kit were 100 and 96 %, respectively. Primary evaluation showed that the new latex agglutination kit was able to quickly diagnose the K99+ bacteria and can be used for the detection of calves colibacillosis.
KeywordsEscherichia coli K99 Latex agglutination test PCR
This research was carried out with the financial support of Vice Chancellor of Research and Technology of Shahid Bahonar University of Kerman. Therefore, the authors would like to appreciate the members of this department for their efforts and acknowledge the personnel in Microbiology Laboratory, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
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