Comparative evaluation of a laboratory-developed real-time PCR assay and RealStar® Adenovirus PCR Kit for quantitative detection of human adenovirus
| العنوان: | Comparative evaluation of a laboratory-developed real-time PCR assay and RealStar® Adenovirus PCR Kit for quantitative detection of human adenovirus |
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| المؤلفون: | Andrew Kai-Chun Cheng, Kwok-Hung Chan, Cyril C. Y. Yip, Vincent C.C. Cheng, Ami M. Y. Fung, Kwok-Yung Yuen, Ho-Yin Lam, Siddharth Sridhar, Jasper Fuk-Woo Chan, Kit-Hang Leung, Samson S. Y. Wong, Bone S.F. Tang |
| المصدر: | Virology Journal, Vol 15, Iss 1, Pp 1-9 (2018) Virology Journal |
| بيانات النشر: | BMC, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, Adenoviridae Infections, 030106 microbiology, RealStar® Adenovirus PCR Kit, Biology, Real-Time Polymerase Chain Reaction, Sensitivity and Specificity, Adenoviral infections, Comparative evaluation, lcsh:Infectious and parasitic diseases, 03 medical and health sciences, 0302 clinical medicine, Virology, Proficiency testing, Humans, lcsh:RC109-216, 030212 general & internal medicine, Detection limit, Adenoviruses, Human, High mortality, Methodology, Human adenovirus, Reproducibility of Results, Viral Load, Infectious Diseases, Real-time polymerase chain reaction, Molecular Diagnostic Techniques, Viral Transport medium, Viral load, Real-time PCR |
| الوصف: | Background Human adenoviruses are common causes of community-acquired respiratory tract and enteric infections. Severe disseminated infections with high mortality rates may be seen in immunocompromised individuals. An accurate and cost-effective quantitative assay is essential not only for laboratory diagnosis of adenoviral infections, but also for monitoring of response to antiviral treatment. The diagnostic performance of an in-house quantitative polymerase chain reaction assay was compared to a commercial system. Methods The analytical sensitivity, specificity, linearity, precision and accuracy of an in-house adenovirus quantitative polymerase chain reaction assay were evaluated against the RealStar® Adenovirus PCR Kit (Altona Diagnostics GmbH, Hamburg, Germany), using 122 clinical specimens and 18 proficiency testing samples. Results Linear regression analysis of the quantitative results by the in-house assay showed the dynamic range from 2.60 to 9 log10 (plasma) and 2.94 to 9 log10 (viral transport medium) copies/mL, with the coefficient of determination (R2) of 0.996 and 0.998, respectively. A dilution series demonstrated the limits of detection and lower limits of quantification for plasma were 2.06 log10 and 2.60 log10 copies/mL and those for viral transport medium were 2.31 log10 and 2.94 log10 copies/mL respectively. The precision of the in-house assay was highly reproducible among runs with coefficients of variance ranging from 0.07 to 3.21% for plasma and 0.17% to 2.11% for viral transport medium. A comparison of 52 matched samples showed an excellent correlation between the quantitative viral loads measured by the in-house assay and the RealStar® Adenovirus PCR Kit (R2 = 0.984), with an average bias of − 0.16 log10 copies/mL. Conclusions The in-house adenovirus assay is a sensitive and reliable assay with lower cost for the detection and quantification of adenoviral DNA when compared to the RealStar® Adenovirus PCR Kit. Electronic supplementary material The online version of this article (10.1186/s12985-018-1059-7) contains supplementary material, which is available to authorized users. |
| اللغة: | English |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1a7321489ad5ba182c8f40bdd9c746e3 http://link.springer.com/article/10.1186/s12985-018-1059-7 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....1a7321489ad5ba182c8f40bdd9c746e3 |
| قاعدة البيانات: | OpenAIRE |
| الوصف غير متاح. |