title: Genotyping of classical swine fever virus using high-resolution melt analysis.
abstract: Discrimination between different field and vaccine strains of classical swine fever virus (CSFV) is crucial for meaningful disease diagnosis and epidemiological investigation. In this study, a rapid method for differentiating vaccine strains and outbreak CSFV isolates by combined RT-PCR and high-resolution melt (HRM) analysis has been developed. The assay is based on PCR amplification of short fragments from the most variable region of CSFVgene E2, followed by HRM analysis of amplicons. Real-Time PCR/HRM for CSFV detection and differentiation analysis has sensitivity comparable to RT-qPCR and genotyping resolution comparable to E2 nucleotide sequencing. This assay in one step enables rapid and sensitive identification and genotype discrimination of CSFV in field samples, and thus will be valuable for CSF outbreak response and disease control.
authors: National Research Institute of Veterinary Virology and Microbiology Russian Academy of Agricultural science, Pokrov, Russia.
authors: Ilya
authors: I
authors: Titov
authors: National Research Institute of Veterinary Virology and Microbiology Russian Academy of Agricultural science, Pokrov, Russia.
authors: Sodnom
authors: S
authors: Tsybanov
authors: National Research Institute of Veterinary Virology and Microbiology Russian Academy of Agricultural science, Pokrov, Russia. Electronic address: Malogolovkin@inbox.ru.
authors: Alexander
authors: A
authors: Malogolovkin
mesh_terms: D000818:Animals; D006691:Classical Swine Fever / Q000175:diagnosis / Q000821:virology; D006692:Classical Swine Fever Virus / Q000145:classification* / Q000235:genetics*; D005838:Genotype; D060005:Genotyping Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013552:Swine; D044366:Transition Temperature
keywords: Classical swine fever virus; E2 gene; Genogroups; Genotyping; HRM; High-resolution melt analysis; RT-qPCR