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Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus.

ABSTRACT:

Background

In this study, we developed a SYBR Green-based real-time PCR assay for the detection of sheep pox virus using a plasmid construct carrying one of the highly conserved genes encoding the virion envelope protein (P32) as a template.

Results

The method was demonstrated to be highly sensitive, allowing a precise SPV DNA quantitation over a range of nine orders of magnitude (from 101 to 109 copies of standard DNA). Then, specimens from SPV suspected sheep were analyzed by conventional gel-based PCR, real-time PCR and sequence analysis.

Conclusion

Comparison between these different techniques revealed that real-time PCR is more sensitive than conventional gel-based PCR, allowing detection low viral titers of SPV in infected sheep.

SUBMITTER: Tian H

PROVIDER: S-EPMC3546957 | biostudies-literature | 2012 Nov

REPOSITORIES: biostudies-literature

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Development of a SYBR green real-time PCR method for rapid detection of sheep pox virus.

Tian Hong H Wu Jingyan J Chen Yan Y Zhang Keshan K Shang Youjun Y Liu Xiangtao X

Virology journal 20121127

<h4>Background</h4>In this study, we developed a SYBR Green-based real-time PCR assay for the detection of sheep pox virus using a plasmid construct carrying one of the highly conserved genes encoding the virion envelope protein (P32) as a template.<h4>Results</h4>The method was demonstrated to be highly sensitive, allowing a precise SPV DNA quantitation over a range of nine orders of magnitude (from 101 to 109 copies of standard DNA). Then, specimens from SPV suspected sheep were analyzed by co ...[more]

PMID: 23186407

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