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Development of a TaqMan Based Real-Time Fluorescent Quantitative PCR Assay for Detection of Porcine Cytomegalovirus in Semen

Authors :
Chen Qiuyong
Wu Xuemin
Yan Shan
Chen Rujing
Long-bai Wang
Wang Chenyan
Lun-jiang Zhou
Che Yongliang
Source :
BioMed Research International, Vol 2020 (2020), BioMed Research International
Publication Year :
2020
Publisher :
Hindawi Limited, 2020.

Abstract

This study described a TaqMan based real-time fluorescent quantitative PCR (qPCR) method to detect porcine cytomegalovirus (PCMV) infection, targeting the conserved region of the DNA polymerase (DPOL) gene. The standard curve showed a linear regression relationship with a coefficient of 0.999 and a slope of y = −3.249x + 38.958 corresponding to the amplification efficiency at 99.8%. The limit of the qPCR method was 51.9 copies/μl. The established qPCR method showed excellent specificity, with no cross-reaction observed with common porcine pathogens. The coefficient of variation for intra-assay and interassay variability ranged up to 1.51% and 2.24%, respectively. PCMV positive signals can be found in semen using this qPCR method, which suggested that we should pay more attention to PCMV contamination in semen in order to eliminate PCMV infection in artificial insemination and xenotransplantation.

Details

Language :
English
ISSN :
23146141 and 23146133
Volume :
2020
Database :
OpenAIRE
Journal :
BioMed Research International
Accession number :
edsair.doi.dedup.....410f042bd2559eb655c214aadaa63fd7