Development of a primer-probe energy transfer real-time PCR assay for improved detection of classical swine fever virus

• Published in: Journal of virological methods Vol. 160; no. 1; pp. 69 - 73
• Main Authors: Liu, Lihong, Xia, Hongyan, Belák, Sándor, Widén, Frederik
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 01.09.2009; Elsevier
• Subjects: Animal and Dairy Science; Animals; Biological and medical sciences; C-strain vaccine; Classical Swine Fever - diagnosis; Classical Swine Fever - virology; Classical swine fever virus; Classical swine fever virus - genetics; Classical swine fever virus - isolation & purification; CSFV; DNA Primers - chemistry; DNA Primers - genetics; Fluorescence Resonance Energy Transfer; Fundamental and applied biological sciences. Psychology; Husdjursvetenskap; Microbiology; Oligonucleotide Probes - chemistry; Oligonucleotide Probes - genetics; Polymerase Chain Reaction - methods; PriProET; Real-time PCR; Reproducibility of Results; Sensitivity and Specificity; Swine; Techniques used in virology; Veterinary Science; Veterinärmedicin; Virology; Virology - methods; C-strain vaccine; PriProET; Real-time PCR; CSFV; Pestivirus; Microbiology; Vaccine strain; Method; Real time; Virology; Virus; Polymerase chain reaction; Flaviviridae; Veterinary; Detection; Hog cholera virus
• ISSN: 0166-0934; 1879-0984; 1879-0984
• DOI: 10.1016/j.jviromet.2009.04.019

Classical swine fever (CSF) is a contagious and devastating disease, causing serious losses in the pig industry worldwide. Vaccination of pigs with the conventional C-strain vaccine has been practised in different regions of the world in order to prevent the disease. In the control programmes of CSF, rapid detection and identification of the causing agent, classical swine fever virus (CSFV) is a crucial step. This study describes a novel real-time PCR assay based on primer-probe energy transfer (PriProET) technology for improved detection of CSFV. The assay is able to detect 20 copies of viral cDNA per reaction, showing a high sensitivity. The specificity has been evaluated by testing 57 pestiviruses, representing all species and unclassified pestiviruses. The assay has been found to be highly reproducible. Following PCR amplification, melting curve analysis allows confirmation of specific amplicons, and differentiation between wild-type CSFV and certain C-strain vaccines. This study provides a new tool for the diagnosis of CSF.