van der Logt J T, Kissing J, Melchers W J
Department of Medical Microbiology, University of Nijmegen, The Netherlands.
J Clin Microbiol. 1994 Aug;32(8):2003-6. doi: 10.1128/jcm.32.8.2003-2006.1994.
To improve the detection of lactate dehydrogenase-elevating virus (LDV), we developed a PCR assay. Primers were selected from ORF7, encoding nucleocapsid protein VP1. No specific amplification was observed with any other common murine virus or with RNAs from the closely related Lelystad virus and equine arteritis virus. In experimentally infected mice, LDV could be detected in plasma in both the acute and the persistent phases. LDV was also detected by the PCR in contaminated pools of Plasmodium berghei parasites which were maintained in mice, both by a direct analysis of the samples and by testing of plasma from mice inoculated with these pools. There was a complete agreement between the results of the PCR assay and the lactate dehydrogenase (LDH) enzyme assay of plasma from the inoculated mice. In contrast to the results of the LDH enzyme assay, no false-positive reactions were obtained in the PCR assay with negative control samples showing visible hemolysis. Storage of plasma samples at room temperature and at 4, -20, and -80 degrees C for up to 8 days did not influence the results of the PCR. These results show that the PCR is a valuable technique which may replace the LDH test as a diagnostic tool.
为了提高乳酸脱氢酶升高病毒（LDV）的检测率，我们开发了一种聚合酶链反应（PCR）检测方法。引物选自编码核衣壳蛋白VP1的开放阅读框7（ORF7）。未观察到与任何其他常见鼠病毒或来自密切相关的莱利斯塔德病毒和马动脉炎病毒的RNA发生特异性扩增。在实验感染的小鼠中，在急性期和持续期的血浆中均可检测到LDV。通过对样本的直接分析以及对接种这些样本的小鼠血浆进行检测，在保存在小鼠体内的受污染伯氏疟原虫池中也通过PCR检测到了LDV。PCR检测结果与接种小鼠血浆的乳酸脱氢酶（LDH）酶检测结果完全一致。与LDH酶检测结果相反，在PCR检测中，对显示可见溶血的阴性对照样本未获得假阳性反应。将血浆样本在室温、4℃、-20℃和-80℃下保存长达8天不影响PCR结果。这些结果表明，PCR是一种有价值的技术，可作为诊断工具替代LDH检测。
