title: Simultaneous detection of major enteric viruses using a combimatrix microarray.
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.
authors.affiliation: Department of Environmental Health and Institute of Health and Environment, School of Public Health, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Ju-Mi
authors.initials: JM
authors.lastname: Kim
authors.forename: Sung Yeon
authors.initials: SY
authors.lastname: Kim
authors.forename: Young Bin
authors.initials: YB
authors.lastname: Park
authors.forename: Hye Jin
authors.initials: HJ
authors.lastname: Kim
authors.forename: Byung Sup
authors.initials: BS
authors.lastname: Min
authors.forename: Jae-Chang
authors.initials: JC
authors.lastname: Cho
authors.forename: Jai Myung
authors.initials: JM
authors.lastname: Yang
authors.forename: You-Hee
authors.initials: YH
authors.lastname: Cho
authors.forename: GwangPyo
authors.initials: G
authors.lastname: Ko
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000302:isolation & purification*; D019200:Astroviridae / Q000235:genetics / Q000302:isolation & purification*; D005759:Gastroenteritis / Q000175:diagnosis / Q000821:virology; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D029322:Norovirus / Q000235:genetics / Q000302:isolation & purification*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D015345:Oligonucleotide Probes