Rimstad E, Ueland K
Department of Microbiology and Immunology, Norwegian College of Veterinary Medicine, Oslo.
J Virol Methods. 1992 Mar;36(3):239-48. doi: 10.1016/0166-0934(92)90055-i.
A specific and sensitive polymerase chain reaction (PCR) procedure for the detection of feline immunodeficiency virus (FIV) in peripheral blood mononuclear cells (PBMC) was developed. PBMC from both blood samples and cultures were digested by proteinase K in a lysis buffer, and after heat inactivation of the proteinase, the resultant material was used in a two step amplification protocol using nested sets of primers. Two independent amplifications, from the gag and pol genes respectively, were performed in each tube. The PCR was positive for six of 14 samples from FIV seropositive adult cats, while all 36 samples from seronegative cats were negative. In comparison with an antigen-capturing ELISA procedure, the PCR detected FIV infection in PBMC cultures on average two days earlier.
开发了一种用于检测外周血单核细胞(PBMC)中猫免疫缺陷病毒(FIV)的特异性和灵敏的聚合酶链反应(PCR)方法。来自血液样本和培养物的PBMC在裂解缓冲液中用蛋白酶K消化,蛋白酶热失活后,所得材料用于使用嵌套引物组的两步扩增方案。在每个管中分别从gag和pol基因进行两次独立扩增。来自FIV血清阳性成年猫的14个样本中有6个PCR呈阳性,而来自血清阴性猫的所有36个样本均为阴性。与抗原捕获ELISA方法相比,PCR平均提前两天检测到PBMC培养物中的FIV感染。