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 bacterium and its toxin. Thus, many trials including microarray-based methods have been applied for rapid, sensitive, and selective detection. However, DNA or protein microarray has been limited to only genotype- or phenotype-based detection. Also, it is difficult to discriminate between CT and heat-labile toxin (LT) from Escherichia coli using commercially available method, because LT is functionally, structurally, and immunologically similar with CT. In the present study, we developed a new microarray, which is composed of two distinct biomolecular-markers: DNA (capture probe from 16S rDNA) and carbohydrate (GM1 as capture probe). This novel double biomolecular marker array is rapid, specific, and simple not only to detect bacteria but also to analyze their pathogenicity through phenotype-based toxin detections. Secreted toxin, CT or LT, 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 and LT. 16S rDNA-derived DNA capture probes can detect bacterium species specifically and estimate which bacterium produced the toxin. Target materials such as chromosomal DNA and toxin can be directly obtained from bacterial cells by one-step preparation. Thus, our newly proposed double biomolecular marker microarray is the efficient and powerful tool to detect bacteria and identify pathogenicity simultaneously.

Detection of toxin producing Vibrio cholerae and Escherichia coli using double biomolecular marker microarray

249th National Meeting (2015)

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Detection of toxin producing Vibrio</i><i> cholerae</i> and <i>Escherichia coli</i> using double biomolecular marker microarray