Kappeler A, Lutz-Wallace C, Sapp T, Sidhu M
Biologics Evaluation Laboratory, Animal Diseases Research Institute, Agriculture and Agri-Food Canada, Nepean, Ontario, Canada.
Biologicals. 1996 Jun;24(2):131-5. doi: 10.1006/biol.1996.0017.
A nested polymerase chain reaction (PCR) assay has been developed for the detection of bovine polyomavirus (BPyV) DNA. The assay has been used to screen commercial lots of fetal bovine serum and modified live veterinary vaccines for the presence of the agent. A PCR product of the expected size was detected after the first round of PCR for eight out of 20 serum lots, but in none of the 14 vaccines tested. The subsequent nested assay revealed that four more serum lots were positive for BPyV DNA, as well as two vaccine lots. When hybridized with a labelled probe, blots of the PCR products from vaccines revealed that in one of the two positive samples a specific product was present after the first PCR at a level not detectable in gel electrophoresis. Nested PCR appears to be a useful tool for the detection of low level contamination with BPyV DNA of products used in, and derived from cell culture.
已开发出一种巢式聚合酶链反应（PCR）检测方法用于检测牛多瘤病毒（BPyV）DNA。该检测方法已用于筛查商业批量的胎牛血清和改良活兽用疫苗中是否存在该病原体。在第一轮PCR后，20个血清批次中有8个检测到预期大小的PCR产物，但在测试的14种疫苗中均未检测到。随后的巢式检测显示，又有4个血清批次以及2个疫苗批次的BPyV DNA呈阳性。当与标记探针杂交时，疫苗PCR产物的印迹显示，在两个阳性样品中的一个中，第一次PCR后存在一种在凝胶电泳中无法检测到水平的特异性产物。巢式PCR似乎是检测用于细胞培养以及源自细胞培养的产品中BPyV DNA低水平污染的有用工具。
