Development and application of TaqMan-MGB real-time quantitative PCR assay for detection of goat pox virus.
- Author:
Zhentao CHENG
1
;
Jun YUE
;
Yongming LI
;
Leren XU
;
Kaigong WANG
;
Bijun ZHOU
;
Junyi CHEN
;
Jun LI
;
Nan JIANG
Author Information
1. Institute of Animal Disease, Guizhou University, Guiyang 550025, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Base Sequence;
Capripoxvirus;
genetics;
isolation & purification;
Goats;
Molecular Sequence Data;
Polymerase Chain Reaction;
methods;
Poxviridae Infections;
diagnosis;
virology;
Sensitivity and Specificity
- From:
Chinese Journal of Biotechnology
2009;25(3):464-472
- CountryChina
- Language:Chinese
-
Abstract:
The complete gene sequences of eight capripoxvirus strains in GenBank were aligned and analyzed with DNAStar software. We selected a size of 64 bp gene fragment that was located in gp064 region of goat pox virus (GPV) genome, and designed a pair of primers and a TaqMan-MGB probe against the gene fragment with Primer Express 2.0 software. Then, the fluorescence quantitative PCR (FQ-PCR) assay was developed and the standard curve of different dilution series was described. We extracted the DNA samples from clinical skin pox, scab and GPV infected materials of artificial challenge animals. The FQ-PCR assay has been performed for all kinds of DNA samples. The results showed that the FQ-PCR assay was sensitive, specific, stable and could be used for clinical diagnosis. This method provided an important tool for rapid diagnosis of goat pox clinically, and for study GPV pathogenesis in the course of disease occurrence, development and convalescence.
