Resequencing microarray for detection of human adenoviruses in patients with community-acquired gastroenteritis: a proof-of-concept study

Woo, Patrick C. Y.; Lau, Susanna K. P.; Lee, Paul; Jiang, Miao; Fung, Ami M. Y.; Choi, Garnet Kwan-Yue; Ellis-Behnke, Rutledge; Yuen, Kwok‐Yung

doi:10.1099/jmm.0.023796-0

Cited by 5 publications

(2 citation statements)

References 13 publications

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“…Therefore, existing HAdV-Ds represent an evolutionary sampling of potential diversity, and proteotype analysis defines the available recombination palette of hypervariable regions for the evolution of new HAdV-Ds. It is also important to note that the evolution of new HAdVs by homologous recombination requires co-infection in the same host cell of at least two unique genotypes from the same viral species36373839404142. The emergence of new HAdV-Ds during the AIDS epidemic suggests a role for persistence of multiple viruses under reduced immune surveillance434445.…”

Section: Discussionmentioning

confidence: 99%

Molecular evolution of human adenoviruses

Robinson

Singh

Lee

et al. 2013

Sci Rep

227

305

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The recent emergence of highly virulent human adenoviruses (HAdVs) with new tissue tropisms underscores the need to determine their ontogeny. Here we report complete high quality genome sequences and analyses for all the previously unsequenced HAdV serotypes (n = 20) within HAdV species D. Analysis of nucleotide sequence variability for these in conjunction with another 40 HAdV prototypes, comprising all seven HAdV species, confirmed the uniquely hypervariable regions within species. The mutation rate among HAdV-Ds was low when compared to other HAdV species. Homologous recombination was identified in at least two of five examined hypervariable regions for every virus, suggesting the evolution of HAdV-Ds has been highly dependent on homologous recombination. Patterns of alternating GC and AT rich motifs correlated well with hypervariable region recombination sites across the HAdV-D genomes, suggesting foci of DNA instability lead to formulaic patterns of homologous recombination and confer agility to adenovirus evolution.

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Section: Discussionmentioning

confidence: 99%

Molecular evolution of human adenoviruses

Robinson

Singh

Lee

et al. 2013

Sci Rep

227

305

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“…In recent years a commercial qPCR assay kit was available for specific detection of HAdV which facilitates the diagnosis and monitoring of HAdV infections in clinical laboratories [ 17 ]. Adenovirus PCR is also included in a number of other commercial multiplex PCR systems for clinical diagnostics, and we have shown that some of the newer technologies such as the resequencing microarray could also be used for the diagnosis of gastroenteritis and conjunctivitis due to HAdV [ 18 , 19 ].…”

Section: Introductionmentioning

confidence: 99%

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

Wong

Yip

Sridhar

et al. 2018

Virol J

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BackgroundHuman 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.MethodsThe 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.ResultsLinear 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.ConclusionsThe 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 materialThe online version of this article (10.1186/s12985-018-1059-7) contains supplementary material, which is available to authorized users.

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Adenoviruses

Allen¹,

Demmler-Harrison²

2018

Principles and Practice of Pediatric Infectious Diseases

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Resequencing microarray for detection of human adenoviruses in patients with community-acquired gastroenteritis: a proof-of-concept study

Cited by 5 publications

References 13 publications

Molecular evolution of human adenoviruses

Molecular evolution of human adenoviruses

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

Adenoviruses

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