Development of a recombinase-aided amplification assay for rapid detection of human norovirus GII.4

• Published in: BMC infectious diseases 2021-03, Vol.21 (1), p.248-248, Article 248
• Main Authors: Qin, Zhiwei, Xue, Liang, Cai, Weicheng, Gao, Junshan, Jiang, Yueting, Yang, Jiale, Liang, Yanhui, Wang, Linping, Zhang, Jumei, Hu, Yongdan, Wu, Qingping
• Format: Article
• Language: English
• Subjects: Adenoviruses; Amplification; Assaying; Base Sequence; Biological assay; Caliciviridae Infections - diagnosis; Caliciviridae Infections - virology; Cross-reaction; Design; Detection; DNA Primers - metabolism; Enzymes; Fluorescence; Foodborne diseases; Gastroenteritis - diagnosis; Gastroenteritis - virology; Genetic aspects; Genomes; Genotype; Genotype & phenotype; Health aspects; Humans; Identification and classification; Infectious diseases; Low income groups; Methods; Norovirus; Norovirus - genetics; Norovirus - isolation & purification; Nucleic Acid Amplification Techniques - methods; Nucleic acids; Pathogens; Polymerase chain reaction; Public health; Public health administration; Rapid diagnostic technique; Real time; Recombinase; Recombinase-aided amplification; Recombinases - metabolism; Reverse transcription; RNA, Viral - chemistry; RNA, Viral - genetics; RNA, Viral - metabolism; Sensitivity and Specificity; Sequence Alignment; Viruses
• ISSN: 1471-2334; 1471-2334
• DOI: 10.1186/s12879-021-05942-x

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. 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). 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