Background: Vibrio cholerae is an important agent of diarrheal diseases in many parts of Asia and Africa. It is an enteric pathogen which 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.

Materials and Methods: Six pair 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 genes. Moreover, sensitivity and specificity of this method were determined.

Results: 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.

Conclusion: Hexaplex PCR Assay could be used to design a kit for cholerae detection.