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

详细信息    查看全文

• 作者：Xing-Juan Zhang ; Hongyan Xia ; Helen Everett ; Olubukola Sosan ; Helen Crooke ; Sá ; ndor Belá ; k ; Frederik Widé ; n ; Hua-Ji Qiu ; Lihong Liu
• 关键词：CSFV ; Hog Cholera Lapinized Virus ; Vaccine ; Validation ; Real-time PCR
• 刊名：Journal of Virological Methods
• 出版年：2010
• 出版时间：September 2010
• 年：2010
• 卷：168
• 期：1-2
• 页码：259-261
• 全文大小：108 K

文摘

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.