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

Sidhu, Jatinder P.S., Ahmed, Warish and Toze, Simon (2013) Sensitive detection of human adenovirus from small volume of primary wastewater samples by quantitative PCR. Journal of Virological Methods, 187 2: 395-400. doi:10.1016/j.jviromet.2012.11.002


Author Sidhu, Jatinder P.S.
Ahmed, Warish
Toze, Simon
Title Sensitive detection of human adenovirus from small volume of primary wastewater samples by quantitative PCR
Journal name Journal of Virological Methods   Check publisher's open access policy
ISSN 0166-0934
1879-0984
Publication date 2013-02
Year available 2012
Sub-type Article (original research)
DOI 10.1016/j.jviromet.2012.11.002
Volume 187
Issue 2
Start page 395
End page 400
Total pages 6
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Collection year 2013
Language eng
Formatted abstract
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 were significantly higher (P < 0.01) when Qiagen Blood and Tissue kit (1.54 × 106 L−1) was used as compared to Mo-Bio 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.
Keyword Wastewater
Adenovirus
Virus concentration
PCR inhibitors
Quantitative PCR
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published online: 23 November 2012.

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
School of Public Health Publications
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 6 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 8 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Wed, 16 Jan 2013, 13:02:57 EST by Geraldine Fitzgerald on behalf of School of Public Health