Development of a reverse-transcription polymerase chain reaction assay with fluorogenic probes to discriminate Korean wild-type and vaccine isolates of Classical swine fever virus.
A 1-step reverse-transcription polymerase chain reaction (RT-PCR) assay using TaqMan minor-groove-binding (MGB) probes was developed to distinguish between vaccine-type and wild-type strains of Classical swine fever virus (CSFV) in Korea. Because attenuated Korean LOM strains have been used in animal vaccination in Korea for some time but CSF remains a serious problem, there was a need for a practical approach to differentiating vaccine and field strains. We examined the fluorescence of 5 vaccine strains, 10 field strains, and 5 mixed samples. Three clusters of the samples could be distinguished: those with only fluorescence of the vaccine-type-specific probe, VIC; those with only fluorescence of the wild-type-specific probe, FAM; and those with both VIC and FAM fluorescence. The RT-PCR assay with fluorogenic probes is sensitive and accurate and is therefore useful for differentiating vaccine and field strains of CSFV in Korea.
Cho
Ho-Seong
HS
Department of Veterinary Pathology, College of Veterinary Medicine, Chonnam National University, Gwangju, Republic of Korea.
Park
Suk-Jun
SJ
Park
Nam-Yong
NY
D000818:Animals; D001483:Base Sequence; D006691:Classical Swine Fever / Q000175:diagnosis*; D006692:Classical Swine Fever Virus / Q000145:classification* / Q000302:isolation & purification; D003937:Diagnosis, Differential; D005453:Fluorescence; D007723:Korea; D011110:Polymorphism, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000662:veterinary*; D012680:Sensitivity and Specificity; D013552:Swine; D014611:Vaccination / Q000662:veterinary