Detection of cholera toxin (ctxAand ctxAB) genes in Vibrio choleraeisolated from clinical and environmental samples in Limbang Sarawak by multiplex polymerase chain reaction (PCR)

Amirah Zakirah, Ja’afar and Elexson, Nillian and Lesley Maurice, Bilung and Grace, Bebey and Diyana, Zakaria and Patrick Guda, Benjamin (2021) Detection of cholera toxin (ctxAand ctxAB) genes in Vibrio choleraeisolated from clinical and environmental samples in Limbang Sarawak by multiplex polymerase chain reaction (PCR). Malaysian Journal of Microbiology, 17 (1). pp. 1-8. ISSN 2231-7538

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Abstract

Aims:Cholera epidemics have been occurred in Malaysia since 1991 till 2003 which can be proved from the records by the Infectious Diseases Division of the Ministry of Health. Moreover, therewere also course of cholera epidemics from the year 1994 to 2003 which had been happenedin Sarawak. Cholera outbreaks in Malaysia mostlycaused by the El Tor O1 Vibrio cholerae serogroup. The aims of this study were to detect the presence of V. choleraein clinical and environmental samples (n=28) from Limbang, Sarawak by collaboration withSarawak Government Hospital and to detect the toxin genes from the isolates.Methodology and results:All the isolates were sub-cultured in alkaline peptone water (APW). The boiled-cell method was used for DNA extraction. The total DNA extracted wasamplified by polymerase chain reaction (PCR). Two types of PCR were used in this studywhich are16SrRNA PCR and multiplex PCR. The results obtained from the study found out that 16 out of 28 (57.14%) samples were confirmed to be V.choleraespecies. Four primers specific for V.choleraewere used in multiplex PCR (O1 type, O139 type, ctxAand ctxAB) to confirm the species type and the toxin genes. All samples shown positivefor V.choleraeO1 serotypeand 100% positive to all genes for the identification of ctxAand ctxABgenes. Conclusion, significance and impact of study:From this study, it showed that multiplex PCR can be used for research purposes in molecular genetics fieldinvolving cholera outbreak.

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