Development and evaluation of recombinase-aided amplification assays incorporating competitive internal controls for detection of human adenovirus serotypes 3 and 7

Virology Journal

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[1] 0000 0000 8803 2373, grid.198530.6, NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, No. 155, Changbai Street, Changping District, 102206, Beijing, China;0000 0001 0266 8918, grid.412017.1, University of South China, College of Public Health, No. 28, West Changsheng Street, 421001, Hengyang, Hunan, China;Hunan Provincial Center for Disease Control and Prevention, No. 450, Furong Street, 410005, Changsha, Hunan, China;0000 0001 0266 8918, grid.412017.1, University of South China, College of Public Health, No. 28, West Changsheng Street, 421001, Hengyang, Hunan, China;Hunan Provincial Center for Disease Control and Prevention, No. 450, Furong Street, 410005, Changsha, Hunan, China;0000 0000 8803 2373, grid.198530.6, NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, No. 155, Changbai Street, Changping District, 102206, Beijing, China;Hunan Provincial Center for Disease Control and Prevention, No. 450, Furong Street, 410005, Changsha, Hunan, China;grid.431010.7, The Third Xiangya Hospital of Central South University, 138 Tongzipo Road, Yuelu District, 410013, Changsha City, Hunan Province, China;

关键词: Pneumonia;    Human adenovirus;    Duplex recombinase aided amplification;    Detection;   

DOI  :  10.1186/s12985-019-1178-9

来源: publisher

【 摘 要 】

BackgroundHuman adenoviruses are a common group of viruses that cause acute infectious diseases. Human adenovirus (HAdV) 3 and HAdV 7 cause major outbreaks of severe pneumonia. A reliable and practical method for HAdV typing in clinical laboratories is lacking. A simple, rapid and accurate molecular typing method for HAdV may facilitate clinical diagnosis and epidemiological control.MethodsWe developed and evaluated duplex real-time recombinase-aided amplification (RAA) assays incorporating competitive internal controls for detection of HAdV 3 and HAdV 7, respectively. The assays were performed in a one-step in a single tube reaction at 39° for 20 min.ResultsThe analytical sensitivities of the duplex RAA assays for HAdV 3 and HAdV 7 were 5.0 and 14.8 copies per reaction, respectively (at 95% probability by probit regression analysis). No cross-reaction was observed with other types of HAdV or other common respiratory viruses. The duplex RAA assays were used to detect 152 previously-defined HAdV-positive samples. These results agreed with those obtained using a published triplex quantitative real-time PCR protocol.ConclusionsWe provide the first report of internally-controlled duplex RAA assays for the detection of HAdV 3 and HAdV 7. These assays effectively reduce the rate of false negative results and may be valuable for detection of HAdV 3 and HAdV 7 in clinical laboratories, especially in resource-poor settings.

【 授权许可】

CC BY