Zhiwei Qin1,2, Liang Xue3, Weicheng Cai1, Junshan Gao1, Yueting Jiang4, Jiale Yang1, Yanhui Liang1, Linping Wang1, Jumei Zhang1, Yongdan Hu5, Qingping Wu6. 1. Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, No. 100, Xianlie Zhong Road, Guangzhou, Guangdong, 510070, People's Republic of China. 2. Faculty Agriculture and Food, Kunming University of Science and Technology, Kunming, 650500, Yunnan, China. 3. Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, No. 100, Xianlie Zhong Road, Guangzhou, Guangdong, 510070, People's Republic of China. xueliang@gdim.cn. 4. Department of Laboratory Medicine, First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, 510120, People's Republic of China. 5. Faculty Agriculture and Food, Kunming University of Science and Technology, Kunming, 650500, Yunnan, China. kunlan881221@126.com. 6. Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, No. 100, Xianlie Zhong Road, Guangzhou, Guangdong, 510070, People's Republic of China. wuqp203@163.com.
Abstract
BACKGROUND: Human noroviruses are one of the main causes of foodborne illnesses and represent a serious public health concern. Rapid and sensitive assays for human norovirus detection are undoubtedly necessary for clinical diagnosis, especially in regions without more sophisticated equipment. METHOD: The rapid reverse transcription recombinase-aided amplification (RT-RAA) is a fast, robust and isothermal nucleic acid detection method based on enzyme reaction. This method can complete the sample detection at 39 °C in 30 min. In this study, we successfully established a rapid reverse transcription recombinase-aided amplification (RT-RAA) assay for the detection of human norovirus GII.4 and applied this assay to clinical samples, as well as comparison with commercial reverse transcription real-time fluorescence quantitative PCR (RT-qPCR). RESULTS: At 95% probability, the detection sensitivity of RT-RAA was 3.425 log10 genomic copies (LGC)/reaction. Moreover, no cross-reaction was observed with other norovirus genogroups and other common foodborne viruses. Stool samples were examined by RT-RAA and reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Compared of RT-qPCR, kappa values for human norovirus detection with RT-RAA were 0.894 (p < 0.001), indicating that both assays were in agreement. CONCLUSION: This RT-RAA assay provides a rapid, specific, and sensitive assay for human norovirus detection and is suitable for clinical testing.
BACKGROUND:Human noroviruses are one of the main causes of foodborne illnesses and represent a serious public health concern. Rapid and sensitive assays for humannorovirus detection are undoubtedly necessary for clinical diagnosis, especially in regions without more sophisticated equipment. METHOD: The rapid reverse transcription recombinase-aided amplification (RT-RAA) is a fast, robust and isothermal nucleic acid detection method based on enzyme reaction. This method can complete the sample detection at 39 °C in 30 min. In this study, we successfully established a rapid reverse transcription recombinase-aided amplification (RT-RAA) assay for the detection of humannorovirus GII.4 and applied this assay to clinical samples, as well as comparison with commercial reverse transcription real-time fluorescence quantitative PCR (RT-qPCR). RESULTS: At 95% probability, the detection sensitivity of RT-RAA was 3.425 log10 genomic copies (LGC)/reaction. Moreover, no cross-reaction was observed with other norovirus genogroups and other common foodborne viruses. Stool samples were examined by RT-RAA and reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Compared of RT-qPCR, kappa values for humannorovirus detection with RT-RAA were 0.894 (p < 0.001), indicating that both assays were in agreement. CONCLUSION: This RT-RAA assay provides a rapid, specific, and sensitive assay for humannorovirus detection and is suitable for clinical testing.
Authors: Nancy P Nenonen; Charles Hannoun; Lennart Svensson; Kjell Torén; Lars-Magnus Andersson; Johan Westin; Tomas Bergström Journal: J Clin Microbiol Date: 2014-04-23 Impact factor: 5.948
Authors: Ozlem Yaren; Kevin M Bradley; Patricia Moussatche; Shuichi Hoshika; Zunyi Yang; Shu Zhu; Stephanie M Karst; Steven A Benner Journal: J Virol Methods Date: 2016-08-18 Impact factor: 2.014