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Development of a real-time quantitative PCR assay for detection of a stable genomic region of BK virus.

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Title: Development of a real-time quantitative PCR assay for detection of a stable genomic region of BK virus.
Authors: Iwaki, Kosuke K Browse this author
Qazi, Suhail H Browse this author
Garcia-Gomez, Jean Browse this author
Zeng, Deanna Browse this author
Matsuda, Yasuhiro Browse this author →KAKEN DB
Matsuda, Kazuko Browse this author
Martinez, Monica E Browse this author
Toyoda, Mieko Browse this author
Kore, Arputharaj Browse this author
Stevens, Wesley T Browse this author
Smogorzewski, Miroslaw Browse this author
Iwaki, Daisuke D Browse this author
Qazi, Yasir Browse this author
Iwaki, Yuichi Browse this author
Issue Date: 29-Oct-2010
Publisher: BioMed Central
Journal Title: Virology journal
Volume: 7
Issue: 1
Start Page: 295
End Page: 304
Publisher DOI: 10.1186/1743-422X-7-295
PMID: 21034442
Abstract: Background: BK virus infections can have clinically significant consequences in immunocompromised individuals. Detection and monitoring of active BK virus infections in certain situations is recommended and therefore PCR assays for detection of BK virus have been developed. The performance of current BK PCR detection assays is limited by the existence of viral polymorphisms, unknown at the time of assay development, resulting in inconsistent detection of BK virus. The objective of this study was to identify a stable region of the BK viral genome for detection by PCR that would be minimally affected by polymorphisms as more sequence data for BK virus becomes available. Results: Employing a combination of techniques, including amino acid and DNA sequence alignment and interspecies analysis, a conserved, stable PCR target region of the BK viral genomic region was identified within the VP2 gene. A real-time quantitative PCR assay was then developed that is specific for BK virus, has an analytical sensitivity of 15 copies/reaction (450 copies/ml) and is highly reproducible (CV ≤ 5.0%). Conclusion: Identifying stable PCR target regions when limited DNA sequence data is available may be possible by combining multiple analysis techniques to elucidate potential functional constraints on genomic regions. Applying this approach to the development of a real-time quantitative PCR assay for BK virus resulted in an accurate method with potential clinical applications and advantages over existing BK assays.
Type: article
Appears in Collections:歯学院・歯学研究院 (Graduate School of Dental Medicine / Faculty of Dental Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 松田 康裕

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