Simultaneous Detection of Major Enteric Viruses Using a Combimatrix Microarray

Ju-Mi Kim; Sung Yeon Kim; Young Bin Park; Hye Jin Kim; Byung Sup Min; Jae-Chang Cho; Jai Myung Yang; You-Hee Cho; GwangPyo Ko

doi:10.1007/s12275-012-2228-9

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Research Support, Non-U.S. Gov't
Simultaneous Detection of Major Enteric Viruses Using a Combimatrix Microarray: Ju-Mi Kim ¹, Sung Yeon Kim ¹, Young Bin Park ¹, Hye Jin Kim ², Byung Sup Min ³, Jae-Chang Cho ², Jai Myung Yang ³, You-Hee Cho ¹, GwangPyo Ko ^1,4; Journal of Microbiology 2012;50(6):970-977.
DOI: https://doi.org/10.1007/s12275-012-2228-9
Published online: October 27, 2012

¹Department of Environmental Health and Institute of Health and Environment, School of Public Health, Seoul National University, Seoul 151-742, Republic of Korea, ²Institute of Environmental Sciences and Department of Environmental Sciences, Hankuk University of Foreign Studies, Yong-In 449-791, Republic of Korea, ³Department of Life Science, Sogang University, Seoul 121-742, Republic of Korea, ⁴Institute of Microbiology, School of Biological Sciences, Seoul National University, Seoul 151-742, Republic of Korea

Corresponding author: GwangPyo Ko , Tel: +82-2-880-2731,

Received: 26 April 2012 • Accepted: 5 July 2012

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Abstract

Various enteric viruses including norovirus, rotavirus, adenovirus, and astrovirus are the major etiological agents of food-borne and water-borne disease outbreaks and frequently cause non-bacterial gastroenteritis worldwide. Sensitive and high-throughput detection methods for these viral pathogens are compulsory for diagnosing viral pathogens and subsequently improving public health. Hence, we developed a sensitive, specific, and high-throughput analytical assay to detect most major enteric viral pathogens using “Combimatrix” platform oligonucleotide probes. In order to detect four different enteric viral pathogens in a sensitive and simultaneous manner, we first developed a multiplex RT-PCR assay targeting partial gene sequences of these viruses with fluorescent labeling for the subsequent microarray. Then, five olignonucleotides specific to each of the four major enteric viruses were selected for the microarray from the oligonulceotide pools targeting the specific genes obtained by multiplex PCR of these viruses. The oligonucleotide microarray was evaluated against stool specimens containing single or mixed viral species. As a result, we demonstrated that the multiplex RT-PCR assay specifically amplified partial sequences of four enteric viruses and the subsequent microarray assay was capable of sensitive and simultaneous detection of those viruses. The developed method could be useful for diagnosing enteric viruses in both clinical and environmental specimens.

Keywords: enteric viruses;microarray;DNA chip

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Simultaneous Detection of Major Enteric Viruses Using a Combimatrix Microarray

J. Microbiol. 2012;50(6):970-977. Published online October 27, 2012

DOI: https://doi.org/10.1007/s12275-012-2228-9

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