Am J Trop Med Hyg 2019 Jun;100(6):1328-1334
Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia.
The diarrheal disease "cholera" is caused by , and is primarily confined to endemic regions, mostly in Africa and Asia. It is punctuated by outbreaks and creates severe challenges to public health. The disease-causing strains are most-often members of serogroups O1 and O139. PCR-based methods allow rapid diagnosis of these pathogens, including the identification of their biotypes. However, this necessitates the selection of specific target sequences to differentiate even the closely related biotypes of . Oligonucleotides for selective amplification of small RNA (sRNA) genes that are specific to these subtypes were designed. The resulting multiplex PCR assay was validated using cultures (i.e., 19 and 22 non- isolates) and spiked stool samples. The validation using cultures and spiked stool suspensions revealed detection limits of 10-100 pg DNA per reaction and 1.5 cells/mL suspension, respectively. The multiplex PCR assay that target sRNA genes for amplification enables the sensitive and specific detection, as well as the differentiation of -O1 classical, O1 El Tor, and O139 biotypes. Most importantly, the assay enables fast and cheaper diagnosis compared with classic culture-based methods.