Sensitive detection of human adenovirus from small volume of primary wastewater samples by quantitative PCR

Jatinder P S Sidhu; Warish Ahmed; S Toze

doi:10.1016/j.jviromet.2012.11.002

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Sensitive detection of human adenovirus from small volume of primary wastewater samples by quantitative PCR

Journal article

Open access

Peer reviewed

Sensitive detection of human adenovirus from small volume of primary wastewater samples by quantitative PCR

Jatinder P S Sidhu, Warish Ahmed and S Toze

Journal of Virological Methods, Vol.187(2), pp.395-400

2013

DOI: https://doi.org/10.1016/j.jviromet.2012.11.002

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Abstract

wastewater

adenovirus

virus concentration

PCR inhibitors

quantitative PCR

An accurate quantitative detection of enteric viruses from the primary wastewater requires, sample concentration followed by extraction of nucleic acid with high purity. A highly efficient and sensitive method was developed for the concentration and quantitative detection of human adenovirus (HAdv) from wastewater samples. The two-step method which combines concentration of virus from 10 mL sample with centrifugal filters followed by extraction and purification of DNA with commercially available nucleic acid extraction kit resulted in high purity DNA for downstream quantitative PCR (qPCR). The results obtained on analytical sensitivities of five commercial nucleic acid extraction kits show that they differ in their ability for DNA yield and purity. Nevertheless, despite variable analytical sensitivities extracted nucleic acid was found to be relatively PCR inhibition free. The genomic copy numbers of HAdv detected from the same concentrated wastewater sample was significantly higher (P less 0.01) when Qiagen Blood and Tissue kit (1.54 × 106 L-1) was used as compared to MoBio PowerSoil kit (5.30 × 105 L-1) which suggests that the nucleic acid extraction kit can influence the sensitivity of qPCR assays. The method developed in this study is simple, rapid, sensitive, and can be applicable for the qPCR detection of adenovirus and other DNA virus in wastewater.

Details

Title: Sensitive detection of human adenovirus from small volume of primary wastewater samples by quantitative PCR
Authors: Jatinder P S Sidhu (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
Warish Ahmed (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
S Toze (Author) - University of Queensland
Publication details: Journal of Virological Methods, Vol.187(2), pp.395-400
Publisher: Elsevier BV
Date published: 2013
DOI: 10.1016/j.jviromet.2012.11.002
ISSN: 0166-0934
Organisation Unit: University of the Sunshine Coast, Queensland
Language: English
Record Identifier: 99450273502621
Output Type: Journal article

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Collaboration types: Domestic collaboration
Web Of Science research areas: Biochemical Research Methods; Biotechnology & Applied Microbiology; Virology

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