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

BACKGROUND: 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 molecul...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6604330/
https://www.ncbi.nlm.nih.gov/pubmed/31262315
http://dx.doi.org/10.1186/s12985-019-1178-9
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author 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
author_facet 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
author_sort Wang, Rui-huan
collection PubMed
description BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
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spelling pubmed-66043302019-07-12 Development and evaluation of recombinase-aided amplification assays incorporating competitive internal controls for detection of human adenovirus serotypes 3 and 7 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 Virol J Methodology BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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. BioMed Central 2019-07-01 /pmc/articles/PMC6604330/ /pubmed/31262315 http://dx.doi.org/10.1186/s12985-019-1178-9 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology
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
Development and evaluation of recombinase-aided amplification assays incorporating competitive internal controls for detection of human adenovirus serotypes 3 and 7
title Development and evaluation of recombinase-aided amplification assays incorporating competitive internal controls for detection of human adenovirus serotypes 3 and 7
title_full Development and evaluation of recombinase-aided amplification assays incorporating competitive internal controls for detection of human adenovirus serotypes 3 and 7
title_fullStr Development and evaluation of recombinase-aided amplification assays incorporating competitive internal controls for detection of human adenovirus serotypes 3 and 7
title_full_unstemmed Development and evaluation of recombinase-aided amplification assays incorporating competitive internal controls for detection of human adenovirus serotypes 3 and 7
title_short Development and evaluation of recombinase-aided amplification assays incorporating competitive internal controls for detection of human adenovirus serotypes 3 and 7
title_sort development and evaluation of recombinase-aided amplification assays incorporating competitive internal controls for detection of human adenovirus serotypes 3 and 7
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6604330/
https://www.ncbi.nlm.nih.gov/pubmed/31262315
http://dx.doi.org/10.1186/s12985-019-1178-9
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