Evaluation of real-time reverse transcription polymerase chain reaction assays for the detection of swine vesicular disease virus

• Published in: Journal of virological methods 2004-03, Vol.116 (2), p.169-176
• Main Authors: Reid, Scott M, Ferris, Nigel P, Hutchings, Geoffrey H, King, Donald P, Alexandersen, Soren
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Biological and medical sciences; Diagnosis; DNA Primers; Enterovirus B, Human - genetics; Enterovirus B, Human - isolation & purification; Fluorogenic RT-PCR; Fundamental and applied biological sciences. Psychology; Geography; Microbiology; Molecular Sequence Data; Real-time RT-PCR; Reverse Transcriptase Polymerase Chain Reaction - methods; Reverse transcription polymerase chain reaction; RNA, Viral - genetics; RNA, Viral - isolation & purification; Swine; Swine Vesicular Disease - diagnosis; Swine vesicular disease virus; Techniques used in virology; Virology
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2003.11.007

Differential detection of swine vesicular disease virus (SVDV) from the other vesicular disease viruses of foot-and-mouth disease (FMD), vesicular stomatitis (VS) and vesivirus is important as the vesicular lesions produced by these viruses are indistinguishable in pigs. Two independent sets of primers and probe, designed from nucleotide sequences within the 5′ untranslated region (UTR) of the SVDV genome, were evaluated in a real-time (5′ nuclease probe-based or fluorogenic) PCR format. Although both primers/probe sets failed to detect one isolate, the assays successfully amplified RNA extracted from epithelial suspensions (ES) and cell culture grown virus preparations from clinical samples representing all currently designated phylogenetic groups of SVDV. Furthermore, no cross-reactivity was demonstrated when these primer/probe sets were tested with RNA prepared from all seven serotypes of FMD virus (FMDV) and from selected isolates of VS virus (VSV), vesivirus and teschoviruses. These assays provide sensitive and rapid alternatives to supplement the routine procedures of ELISA and virus isolation for SVDV diagnosis. The two independent sets of primers/probe can be used routinely while only one of the primers/probe sets would typically be used in SVDV diagnosis during an outbreak.