Cholera is the life-threatening diarrheal disease which is one of major world-wide public health problems. The microbial protein toxin of pathogenic Vibrio cholerae, cholera toxin (CT), is a major causative agent of cholera. Cholera transmission primarily arises from the contaminated food or water. The most significant method for the prevention and surveillance is fast detection and identification of both pathogenic V. cholerae and its toxin. Thus, many trials including microarray-based methods have been applied for rapid, sensitive, and selective detection. However, DNA or protein microarray which respectively aims to detect the cholera toxin genes (ctxAB) or the CT have been limited to only genotype- or phenotype-based detection. In the present study, we developed a new microarray, which is composed of two distinct biomolecular-markers: DNA (capture probes from 16S rDNA and ctxAB), and carbohydrate (GM1). This novel double biomolecular marker array is rapid, specific, and simple not only to detect V. cholerae, but also to analyze their pathogenicity through both genotype- and phenotype-based CT detections. 16S rDNA-derived DNAcapture probes can specifically detect V. cholerae and ctxAB-derived DNA capture probes can check whether the detected V. cholerae have potential pathogenicity. Secreted CT from pathogenic V. cholerae also can be detected by GM1 as carbohydrate capture probe which is cell membrane receptor of the host and has high affinity and specificity with CT. Target materials such as the chromosomal DNA and CT of our microarray can be directly obtained from V. cholerae cells by one-step preparation. Thus, our newly proposed double biomolecular marker microarray, DNA-carbohydrate microarray, is the efficient and powerful tool to detect V. cholerae and simultaneously identify its pathogenicity.

Double biomolecular marker microarray for fast and specific detection of pathogenic <i>Vibrio cholerae</i>

247th National Meeting (2014)

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Double biomolecular marker microarray for fast and specific detection of pathogenic Vibrio cholerae