Besselsen D G, Besch-Williford C L, Pintel D J, Franklin C L, Hook R R, Riley L K
Department of Veterinary Pathology, University of Missouri, Columbia 65211, USA.
J Clin Microbiol. 1995 Jul;33(7):1699-703. doi: 10.1128/jcm.33.7.1699-1703.1995.
H-1 virus and Kilham rat virus (KRV) are autonomous parvoviruses which generally cause subclinical infections in rats and can cause persistent infections in cell cultures. In this study, primer sets specific for either H-1 or KRV were designed on the basis of DNA sequence comparisons of the rodent parvoviruses. The specificities of the H-1 and KRV-specific primer sets were determined by testing viral preparations of seven different parvoviruses and nine other viruses known to infect rodents. The H-1-specific PCR assay amplified the expected 254-bp product only in the presence of H-1 viral DNA and was able to detect as little as 100 fg of H-1 viral DNA. The KRV-specific PCR assay generated the expected 281-bp product only when KRV viral DNA was used as the template and was able to detect as little as 10 pg of KRV viral DNA. Each assay was able to detect its respective virus in tissues from rats experimentally infected with H-1 or KRV. In contrast, no product was amplified by either assay with tissues from mock-infected rats. Our findings indicate that these PCR assays provide rapid, specific, and sensitive methods for the detection of H-1 or KRV infection in rats and cell culture systems.
H-1病毒和基尔汉姆大鼠病毒(KRV)是自主性细小病毒,通常在大鼠中引起亚临床感染,并可在细胞培养物中引起持续性感染。在本研究中,根据啮齿动物细小病毒的DNA序列比较,设计了针对H-1或KRV的引物组。通过检测七种不同细小病毒和已知感染啮齿动物的其他九种病毒的病毒制剂,确定了H-1和KRV特异性引物组的特异性。H-1特异性PCR检测仅在存在H-1病毒DNA时扩增出预期的254 bp产物,并且能够检测低至100 fg的H-1病毒DNA。KRV特异性PCR检测仅在使用KRV病毒DNA作为模板时产生预期的281 bp产物,并且能够检测低至10 pg的KRV病毒DNA。每种检测方法都能够在实验感染H-1或KRV的大鼠组织中检测到各自的病毒。相比之下,用 mock 感染大鼠的组织进行的任何一种检测均未扩增出产物。我们的研究结果表明,这些PCR检测方法为检测大鼠和细胞培养系统中的H-1或KRV感染提供了快速、特异和灵敏的方法。