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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

Zhang, Le, Xue, Liang, Gao, Junshan, Cai, Weicheng, Jiang, Yueting, Zuo, Yueting, Liao, Yingyin, Qin, Zhiwei, Wu, Haoming, Cheng, Tong, Luo, Xueting, Wu, Qingping, Wu, Kegang, Zhang, Jumei
Food control 2020 v.111 pp. 107032
Norovirus, RNA-directed DNA polymerase, detection limit, foodborne illness, gastroenteritis, markets, oysters, proteinases, quantitative polymerase chain reaction, reverse transcriptase polymerase chain reaction
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 × 10³ 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.