Rapid Detection of Vibrio Cholerae Using Hexaplex PCR Assay

Vibrio cholerae is an important agent of diarrheal disease in many parts of Asia and Africa. It is an enteric pathogen which can produce a global pandemic of the disease. Rapid، sensitive and specific measurement of V. cholerae is an interest for many clinical laboratories. The aim of this study was to design and develop a hexaplex PCR assay for rapid detection of V. cholerae. Six pairs of primers were designed for specific amplification of the virulence and regulatory genes for Vibrio cholerae O1: cholera toxin enzymatic subunit A (ctxA) and B (ctxB)، zonula occludens toxin (zot)، accessory cholerae enterotoxin (ace)، toxin- coregulated pilus (tcp) and outer membrane protein (ompW). Hexaplex PCR carried out using six primers and standard gene. Moreover، sensitivity and specificity of this method were determined. Hexaplex PCR showed the presence of the virulence and regulatory genes for Vibrio cholerae O1 in the standard sample. In addition، specificity was qualified and sensitivity determined 100 cfu/ml in this method. Hexaplex PCR Assay could be used to design a kit for cholerae detection.