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

Human 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 me...

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Bibliographic Details
Published in:	Virology journal 2019-07, Vol.16 (1), p.86-86, Article 86
Main Authors:	Wang, Rui-Huan, Zhang, Hong, Zhang, Yi, Li, Xin-Na, Shen, Xin-Xin, Qi, Ju-Ju, Fan, Guo-Hao, Xiang, Xing-Yu, Zhan, Zhi-Fei, Chen, Zi-Wei, Ma, Xue-Jun
Format:	Article
Language:	eng
Subjects:	Adenoviruses Adenoviruses, Human - genetics Adenoviruses, Human - isolation & purification Bioassay Biomedical laboratories Communicable diseases Competition Cross-reaction Deoxyribonucleic acid Detection DNA Duplex recombinase aided amplification Epidemics Epidemiology Human adenovirus Humans Infections Infectious diseases Laboratories Methodology Nucleic Acid Amplification Techniques - methods Nucleic Acid Amplification Techniques - standards Outbreaks Pediatrics Plasmids Pneumonia Polymerase chain reaction Real-Time Polymerase Chain Reaction - methods Real-Time Polymerase Chain Reaction - standards Recombinase Recombinases - genetics Regression analysis Respiratory syncytial virus Respiratory Tract Infections - epidemiology Sensitivity and Specificity Serogroup Serotypes Temperature Typing Viral proteins Viruses
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Summary:	Human 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. We 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. The 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. We 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.
ISSN:	1743-422X 1743-422X