三联RT-ddPCR法在双壳贝类中同时检测人类星状病毒和萨波病毒的应用研究。

IF 1.9 2区 农林科学 Q3 FOOD SCIENCE & TECHNOLOGY
Maolin Wei, Jinfeng Wang, Yan Wang, Libing Liu, Xiangdong Xu, Jianchang Wang
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引用次数: 0

摘要

人类星状病毒(HAstV)和萨波病毒(SaV)是与人类急性胃肠炎相关的重要病原体。本研究建立了以MS2噬菌体为过程控制病毒的三重逆转录-微滴数字聚合酶链式反应(RT-ddPCR)方法,用于同时定量HAstV和SaV。采用240个双壳类样品,包括Ostreidae (n = 43)、Ruditapes philippinarum (n = 84)、Sinonovacula constricta (n = 27)、Scapharca subcrenatata (n = 23)和Pectinidae (n = 63) 5种贝类。结果表明,所建立的RT-ddPCR检测方法具有良好的专有性,对HAstV、SaV和MS2 RNA的检出限分别为5.15拷贝/反应、7.71拷贝/反应和6.13拷贝/反应。病毒筛查结果显示,1.25%(3/240)的样本中存在HAstV,最大载量为22140拷贝/ 2g,而SaV的患病率更高,为13.33%(32/240),峰值浓度为68700拷贝/ 2g。不同种类双壳贝类的检出率不同;SaV检出率最高的是ostreidae,为20.93%(9/43),其次是菲律宾Ruditapes (14.29%) (12/84), Scapharca subcrenatas (13.04%) (3/23), pectinidae (11.11%) (7/63), Sinonovacula constrictas(3.70%)。HAstV仅在菲律宾褐皮菌和果胶菌中检出,检出率分别为1.19%(1/84)和3.17%(2/63)。此外,在单个果胶科样品中均检测到HAstV和SaV(0.42%, 1/240)。本研究建立的三重RT-ddPCR检测方法可靠、准确、灵敏度高,为双壳贝类中HAstV和SaV的定量检测提供了有效的技术支持。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Application Research on the Triplex RT-ddPCR Assay for the Simultaneous Detection of Human Astrovirus and Sapovirus in Bivalve Shellfish.

Human astrovirus (HAstV) and sapovirus (SaV) are significant pathogens associated with acute gastroenteritis in humans. This study established a triple reverse transcription-droplet digital polymerase chain reaction (PCR) (RT-ddPCR) assay incorporating MS2 phage as a process control virus for the simultaneous quantification of HAstV and SaV. The assay was validated using 240 bivalve samples, comprising five shellfish species: Ostreidae (n = 43), Ruditapes philippinarum (n = 84), Sinonovacula constricta (n = 27), Scapharca subcrenata (n = 23), and Pectinidae (n = 63). The results indicated that the developed RT-ddPCR assay had a good exclusivity, with detection limits of 5.15 copies/reaction for HAstV, 7.71 copies/reaction for SaV, and 6.13 copies/reaction for MS2 RNA. Viral screening revealed HAstV in 1.25% (3/240) of samples, with a maximum load of 22,140 copies/2 g, while SaV exhibited a higher prevalence of 13.33% (32/240) and a peak concentration of 68,700 copies/2 g. Different species of bivalve shellfish exhibited varying detection rates; the highest SaV detection rate was found in Ostreidaes at 20.93% (9/43), followed by Ruditapes philippinarums at 14.29% (12/84), Scapharca subcrenatas at 13.04% (3/23), Pectinidaes at 11.11% (7/63), and Sinonovacula constrictas at 3.70% (1/27). HAstV was only detected in R. philippinarums and Pectinidaes, with detection rates of 1.19% (1/84) and 3.17% (2/63), respectively. Additionally, both HAstV and SaV were detected in a single Pectinidae sample (0.42%, 1/240). The triple RT-ddPCR assay developed in this study is reliable, accurate, and highly sensitive, providing effective technical support for the quantitative detection of HAstV and SaV in bivalve shellfish.

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来源期刊
Foodborne pathogens and disease
Foodborne pathogens and disease 医学-食品科技
CiteScore
5.30
自引率
3.60%
发文量
80
审稿时长
1 months
期刊介绍: Foodborne Pathogens and Disease is one of the most inclusive scientific publications on the many disciplines that contribute to food safety. Spanning an array of issues from "farm-to-fork," the Journal bridges the gap between science and policy to reduce the burden of foodborne illness worldwide. Foodborne Pathogens and Disease coverage includes: Agroterrorism Safety of organically grown and genetically modified foods Emerging pathogens Emergence of drug resistance Methods and technology for rapid and accurate detection Strategies to destroy or control foodborne pathogens Novel strategies for the prevention and control of plant and animal diseases that impact food safety Biosecurity issues and the implications of new regulatory guidelines Impact of changing lifestyles and consumer demands on food safety.
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