Development of a high-efficient concentrated pretreatment method for noroviruses detection in independent oysters:An extension of the ISO/TS 15216-2:2013 standard method

被引：13

作者：

Zhang, Le ^{[1
,2
]}

Xue, Liang ^{[2
]}

Gao, Junshan ^{[2
]}

Cai, Weicheng ^{[2
]}

Jiang, Yueting ^{[3
]}

Zuo, Yueting ^{[2
]}

Liao, Yingyin ^{[2
]}

Qin, Zhiwei ^{[2
]}

Wu, Haoming ^{[2
]}

Cheng, Tong ^{[2
]}

Luo, Xueting ^{[2
]}

Wu, Qingping ^{[2
]}

Wu, Kegang ^{[1
]}

Zhang, Jumei ^{[2
]}

机构：

[1] Guangdong Univ Technol, Sch Chem Engn & Light Ind, Guangzhou 510070, Peoples R China

[2] Guangdong Open Lab Appl Microbiol, Guangdong Inst Microbiol Guangdong Acad Sci, Sci State Key Lab Appl Microbiol Southern China, Guangdong Prov Key Lab Microbial Culture Collect, Guangzhou 510070, Peoples R China

[3] Guangzhou Med Univ, Dept Lab Med, Affiliated Hosp 1, Guangzhou 510120, Peoples R China

来源：

FOOD CONTROL | 2020年 / 111卷

基金：

中国国家自然科学基金;

关键词：

Norovirus; Oyster; Virus detection; Protease K; REVERSE-TRANSCRIPTION-PCR; HEPATITIS-A VIRUS; SHELLFISH; QUANTIFICATION; TRANSMISSION; RECOVERY;

D O I：

10.1016/j.foodcont.2019.107032

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

Noroviruses are the primary cause of gastroenteritis and foodborne diseases, affecting millions of individuals annually worldwide. Oysters are frequently associated with norovirus outbreaks. Hence, inexpensive and simple norovirus detection methods are important to detect and contain foodborne illness outbreaks. The objective of this study is to develop a high-efficient concentrated pretreatment method for reverse transcriptase quantitative real-time polymerase chain reaction (RT-qPCR) detection. Based on the existing ISO/TS 15216-2:2013 standard (ISO/TS 15216-2., 2013), four methods are compared for recovery of norovirus from spiked digestive tissue of oysters. A method is found to be the most efficient based on protease K method of increasing buffer volume and PEG precipitation method. The recovery rate and amplification efficiency approached 11.07 +/- 0.09% and 124.12 +/- 5.99%, respectively, being 7-fold that of the original ISO/TS 15216-2:2013 method. This method serves as a rapid (1.5h) sample concentration tool with a limit of detection as low as 7.05 x 10(3) copies. Thirty-eight oyster samples from an aquatic products market in Guangzhou are tested using this method, and noroviruses are detected in 13 samples. This method is an effective extension of the existing ISO/TS 15216-2:2013 standard and it is potentially applicable for detecting norovirus contamination in oysters.

引用

页数：5

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