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用于检测艾奇病毒的逆转录-聚合酶链反应的比较分析

Comparative analysis of reverse-transcription-polymerase chain reaction for Aichivirus detection.

作者信息

Kim Mi-Gyeong, Yoo Boeun, Min A Young, Seo Doo Won, Choi Changsun, Kim Seung Hwan, Kim Soon Han

机构信息

Food Microbiology Division, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety, 187 Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do 28159 Republic of Korea.

School of Food Science and Technology, Chung-Ang University, Ansung, 456-756 South Korea.

出版信息

Food Sci Biotechnol. 2024 Mar 13;33(12):2807-2814. doi: 10.1007/s10068-024-01537-9. eCollection 2024 Sep.

Abstract

(AiV-A), a member of the genus of the family Picornaviridae, was first reported in stool samples of patients with non-bacterial gastroenteritis in Aichi Prefecture, Japan, in 1989. AiV has been reported from in various aquatic environments, such as surface water and sewage, can be transmitted via the fecal-oral route through contaminated water. As AiV is known to acute gastroenteritis worldwide, developing methods for AiV detection from contaminated environments and food is required. In the present study, we established an effective polymerase chain reaction (PCR) method to detect AiV. Various real-time reverse transcription (RT)-PCR and conventional PCR methods for AiV detection were compared, and the limit of detection was confirmed by comparing the sensitivity at varied primer concentrations and PCR conditions. The final detection limits were 10 copy/μL in conventional PCR, and 10 copy/μL in the real-time RT-PCR. The optimized method used in this study might aid in detecting AiV contamination.

摘要

戊型肝炎病毒A(AiV - A)是小RNA病毒科属的成员,于1989年在日本爱知县非细菌性肠胃炎患者的粪便样本中首次被报道。AiV已在各种水生环境中被发现,如地表水和污水,可通过受污染的水经粪口途径传播。由于AiV在全球范围内都与急性肠胃炎有关,因此需要开发从受污染环境和食物中检测AiV的方法。在本研究中,我们建立了一种有效的聚合酶链反应(PCR)方法来检测AiV。比较了各种用于检测AiV的实时逆转录(RT)-PCR和传统PCR方法,并通过比较不同引物浓度和PCR条件下的灵敏度来确定检测限。传统PCR的最终检测限为10拷贝/μL,实时RT-PCR的最终检测限为10拷贝/μL。本研究中使用的优化方法可能有助于检测AiV污染。

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