Evaluation of a primer-probe energy transfer real-time PCR assay for detection of classical swine fever virus.
This study describes evaluation of a real-time PCR assay based on primer-probe energy transfer (PriProET) technology for detection of classical swine fever virus (CSFV). The PriProET technology allows melting curve analysis following PCR amplification and thus provides a higher specificity. The assay was compared with a TaqMan assay by testing a total of 203 samples including 175 clinical specimens and 28 batches of Hog Cholera Lapinized Virus (HCLV) vaccine. The two assays gave the same results for 184 (91%) samples. Compared with the TaqMan assay, 19 additional samples were found to be positive for CSFV using the PriProET assay. In an RNA mixture of both wild type CSFV and C-strain vaccine, the melting curves displayed only one curve: either a wild type-like or a vaccine-like depending on the dominating RNA. The PriProET assay can be a routine molecular tool or a confirmative tool for diagnosis of classical swine fever (CSF), especially in the case of samples that yield an inconclusive result by the TaqMan assay.
Zhang
Xing-Juan
XJ
Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001 Harbin, China.
Xia
Hongyan
H
Everett
Helen
H
Sosan
Olubukola
O
Crooke
Helen
H
Belák
Sándor
S
Widén
Frederik
F
Qiu
Hua-Ji
HJ
Liu
Lihong
L
D000818:Animals; D006691:Classical Swine Fever / Q000175:diagnosis* / Q000821:virology; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics / Q000378:metabolism*; D004735:Energy Transfer; D015345:Oligonucleotide Probes / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine; D044366:Transition Temperature; D014773:Virology / Q000379:methods*
D017931:DNA Primers; D015345:Oligonucleotide Probes