Boerman R H, Arnoldus E P, Raap A K, Bloem B R, Verhey M, van Gemert G, Peters A C, van der Ploeg M
Department of Neurology, University of Leiden, The Netherlands.
J Virol Methods. 1989 Aug;25(2):189-97. doi: 10.1016/0166-0934(89)90032-3.
A technique is described for the detection of HSV-DNA in very small volumes (5-10 microliters) of cerebrospinal fluid (CSF). The method was evaluated in CSF samples of 4-6-week-old mice inoculated with HSV-1 via the corneal route. The sensitivity of the PCR assay was compared with results of spin-amplified viral culture with immunofluorescent visualization (SAC/IF), routine viral culture (RVC) and radioactive dot-blot hybridization (DBA) in CSF samples obtained from other mice. The results show the PCR to be superior over the other techniques: infectious virus or viral DNA in CSF was demonstrated by PCR, SAC/IF, RVC and DBA in 68, 55, 20 and 2.5%, respectively. These results show the feasibility of the PCR for a rapid, non-invasive diagnosis of human HSV-encephalitis.
本文描述了一种用于检测极少量(5 - 10微升)脑脊液(CSF)中单纯疱疹病毒DNA(HSV - DNA)的技术。该方法在经角膜途径接种HSV - 1的4 - 6周龄小鼠的脑脊液样本中进行了评估。将聚合酶链反应(PCR)检测的灵敏度与从其他小鼠获得的脑脊液样本中采用免疫荧光可视化的旋转扩增病毒培养(SAC/IF)、常规病毒培养(RVC)和放射性斑点杂交(DBA)的结果进行了比较。结果显示PCR优于其他技术:脑脊液中的感染性病毒或病毒DNA通过PCR、SAC/IF、RVC和DBA检测出的比例分别为68%、55%、20%和2.5%。这些结果表明PCR用于快速、非侵入性诊断人类HSV脑炎具有可行性。