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Development of a TaqMan Based Real-Time Fluorescent Quantitative PCR Assay for Detection of Porcine Cytomegalovirus in Semen
- 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.
- Subjects :
- 0301 basic medicine
Male
Article Subject
DNA polymerase
Swine
Coefficient of variation
030106 microbiology
Cytomegalovirus
Semen
DNA-Directed DNA Polymerase
Real-Time Polymerase Chain Reaction
Viral Zoonoses
General Biochemistry, Genetics and Molecular Biology
03 medical and health sciences
TaqMan
Animals
Porcine cytomegalovirus
Swine Diseases
General Immunology and Microbiology
biology
General Medicine
Fluorescence
Molecular biology
Standard curve
030104 developmental biology
Real-time polymerase chain reaction
biology.protein
Medicine
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 23146141 and 23146133
- Volume :
- 2020
- Database :
- OpenAIRE
- Journal :
- BioMed Research International
- Accession number :
- edsair.doi.dedup.....410f042bd2559eb655c214aadaa63fd7