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Comparative evaluation of a laboratory-developed real-time PCR assay and RealStar® Adenovirus PCR Kit for quantitative detection of human adenovirus

Authors :
Samson S. Y. Wong
Cyril C. Y. Yip
Siddharth Sridhar
Kit-Hang Leung
Andrew K. W. Cheng
Ami M. Y. Fung
Ho-Yin Lam
Kwok-Hung Chan
Jasper F. W. Chan
Vincent C. C. Cheng
Bone S. F. Tang
Kwok-Yung Yuen
Source :
Virology Journal, Vol 15, Iss 1, Pp 1-9 (2018)
Publication Year :
2018
Publisher :
BMC, 2018.

Abstract

Abstract 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.

Details

Language :
English
ISSN :
1743422X
Volume :
15
Issue :
1
Database :
Directory of Open Access Journals
Journal :
Virology Journal
Publication Type :
Academic Journal
Accession number :
edsdoj.8e09c7e80a0b4b4cbcc2b5de51fb8617
Document Type :
article
Full Text :
https://doi.org/10.1186/s12985-018-1059-7