Liu S T, Li S N, Wang D C, Chang S F, Chiang S C, Ho W C, Chang Y S, Lai S S
Department of Microbiology and Immunology, Chang-Gung Medical College, Taoyuan, Taiwan, R.O.C.
J Virol Methods. 1991 Nov-Dec;35(2):227-36. doi: 10.1016/0166-0934(91)90138-p.
A rapid method for the detection of hog cholera virus (HCV) in infected tissues, using polymerase chain reaction (PCR) was developed. Total RNA isolated from HCV-infected tissues was reverse transcribed with AMV reverse transcriptase and the resulting complementary DNA was amplified by Taq DNA polymerase in the presence of two HCV-specific primers. The amplified DNA fragment was detected by agarose gel electrophoresis. The sensitivity of this method was at 10(4) TCID50 of HCV. The sensitivity increased approximately 1000-fold when the DNA was reamplified with a set of nested primers. DNA sequencing analysis of the PCR products revealed that the HCV sequence amplified from a local field isolate was highly homologous to the HCV Alfort strain. This method may be useful for pathological and epidemiological studies of HCV in pigs.
开发了一种利用聚合酶链反应（PCR）快速检测感染组织中猪瘟病毒（HCV）的方法。从感染HCV的组织中分离出的总RNA用禽成髓细胞瘤病毒（AMV）逆转录酶进行逆转录，所得互补DNA在两种HCV特异性引物存在的情况下由Taq DNA聚合酶进行扩增。扩增的DNA片段通过琼脂糖凝胶电泳进行检测。该方法的灵敏度为10(4) 半数组织培养感染剂量（TCID50）的HCV。当用一组巢式引物对DNA进行再次扩增时，灵敏度提高了约1000倍。对PCR产物的DNA测序分析表明，从本地田间分离株扩增出的HCV序列与HCV阿尔福特株高度同源。该方法可能对猪HCV的病理学和流行病学研究有用。
