Harding M J, Prud'homme I, Gradil C M, Heckert R A, Riva J, McLaurin R, Dulac G C, Vydelingum S
Animal Diseases Research Institute, Ontario, Canada.
J Vet Diagn Invest. 1996 Oct;8(4):414-9. doi: 10.1177/104063879600800402.
A blind panel was tested in a diagnostic evaluation of a reverse transcription (RT) polymerase chain reaction (PCR) method for detecting hog cholera virus (HCV) from pig tissues. The capability of the RT-PCR test to discriminate between HCV and related pestiviruses, bovine viral diarrhea virus (BVDV), and those viruses causing similar diseases in swine, including African swine fever virus (ASFV) and pseudorabies virus (PRV), was also considered. Nucleic acid extraction involved either kit-based or conventional phenol:chloroform:isoamyl alcohol methods. A single-round PCR assay, using primers that hybridize to the conserved p120 nonstructural gene region, was 82.5% sensitive (n = 17) and 100% specific (n = 18) in the detection of the presence of HCV RNA. However, the sensitivity was increased to 100% following a second PCR test. In all, 4 HCV, 7 BVDV, 2 ASFV, and 1 PRV isolates were studied. Novel nucleic acid sequences were generated for 9 HCV strains. Analysis of a portion of the p120 region using these methods was suitable for HCV isolate characterization.
在一项诊断评估中，对一种用于从猪组织中检测猪瘟病毒（HCV）的逆转录（RT）聚合酶链反应（PCR）方法进行了盲法检测。同时还考虑了RT-PCR检测区分HCV与相关瘟病毒、牛病毒性腹泻病毒（BVDV）以及在猪中引起类似疾病的病毒（包括非洲猪瘟病毒（ASFV）和伪狂犬病病毒（PRV））的能力。核酸提取采用基于试剂盒的方法或传统的苯酚：氯仿：异戊醇方法。使用与保守的p120非结构基因区域杂交的引物进行的单轮PCR检测，在检测HCV RNA存在时的灵敏度为82.5%（n = 17），特异性为100%（n = 18）。然而，在进行第二次PCR检测后，灵敏度提高到了100%。总共研究了4株HCV、7株BVDV、2株ASFV和1株PRV分离株。为9株HCV毒株生成了新的核酸序列。使用这些方法对p120区域的一部分进行分析适用于HCV分离株的鉴定。
