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KMID : 0545120180280020210
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Journal of Microbiology and Biotechnology
2018 Volume.28 No. 2 p.210 ~ p.217
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Simultaneous Detection of Four Foodborne Viruses in Food Samples Using a One-Step Multiplex Reverse Transcription PCR
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Lee Shin-Young
Kim Mi-Ju
Kim Hyun-Joong
Jeong Kwang-Cheol Casey
Kim Hae-Yeong
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Abstract
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A one-step multiplex reverse transcription PCR (RT-PCR) method comprising six primer sets (for the detection of norovirus GI and GII, hepatitis A virus, rotavirus, and astrovirus) was developed to simultaneously detect four kinds of pathogenic viruses. The size of the PCR products for norovirus GI and GII, hepatitis A virus (VP3/VP1 and P2A regions), rotavirus, and astrovirus were 330, 164, 244, 198, 629, and 449 bp, respectively. The RT-PCR with the six primer sets showed specificity for the pathogenic viruses. The detection limit of the developed multiplex RT-PCR, as evaluated using serially diluted viral RNAs, was comparable to that of one-step single RT-PCR. Moreover, this multiplex RT-PCR was evaluated using food samples such as water, oysters, lettuce, and vegetable product. These food samples were artificially spiked with the four kinds of viruses in diverse combinations, and the spiked viruses in all food samples were detected successfully.
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KEYWORD
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One-step multiplex reverse transcription PCR, simultaneous detection, norovirus, hepatitis A virus, rotavirus, astrovirus
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