Simultaneous Detection of Major Enteric Viruses Using a Combimatrix Microarray

• Published in: The journal of microbiology Vol. 50; no. 6; pp. 970 - 977
• Main Authors: Kim, J.M., Seoul National University, Seoul, Republic of Korea, Kim, S.Y., Seoul National University, Seoul, Republic of Korea, Park, Y.B., Seoul National University, Seoul, Republic of Korea, Kim, H.J., Hankuk University of Foreign Studies, Yongin, Republic of Korea, Min, B.S., Sogang University, Seoul, Republic of Korea, Cho, J.C., Hankuk University of Foreign Studies, Yongin, Republic of Korea, Yang, J.M., Sogang University, Seoul, Republic of Korea, Cho, Y.H., Seoul National University, Seoul, Republic of Korea, Ko, G.P., Seoul National University, Seoul, Republic of Korea
• Format: Journal Article
• Language: English
• Published: Heidelberg The Microbiological Society of Korea 01.12.2012; Springer Nature B.V; 한국미생물학회
• Subjects: Adenoviridae - genetics; Adenoviridae - isolation & purification; Adenovirus; Adenoviruses; ADN; Assaying; Astroviridae - genetics; Astroviridae - isolation & purification; Astrovirus; Biomedical and Life Sciences; Detection; Disease outbreaks; DNA; DNA chip; DNA microarrays; enteric viruses; Environmental science; Epidemics; Etiology; feces; Fluorescence; fluorescent labeling; Gastroenteritis; Gastroenteritis - diagnosis; Gastroenteritis - virology; Gene sequencing; Genes; Humans; Hybridization; Labeling; Life Sciences; microarray; microarray technology; Microbiology; Multiplex Polymerase Chain Reaction; Multiplexing; Norovirus; Norovirus - genetics; Norovirus - isolation & purification; Nucleotide sequence; nucleotide sequences; Oligonucleotide Array Sequence Analysis - methods; Oligonucleotide Probes; Oligonucleotides; Ozone; Pathogens; PCR; Polymerase chain reaction; Public health; Reverse Transcriptase Polymerase Chain Reaction; Rotavirus; Rotavirus - genetics; Rotavirus - isolation & purification; Viruses; Waterborne diseases; 생물학; South Korea; enteric viruses; microarray; DNA chip
• ISSN: 1225-8873; 1976-3794; 1976-3794
• DOI: 10.1007/s12275-012-2228-9

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.