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首页> 外文期刊>Journal of Clinical Microbiology >Enzymatic amplification of lactate dehydrogenase-elevating virus.
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Enzymatic amplification of lactate dehydrogenase-elevating virus.

机译:酶促扩增乳酸脱氢酶的病毒。

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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.
机译:为了提高对乳酸脱氢酶升高病毒(LDV)的检测,我们开发了一种PCR检测方法。引物选自ORF7,其编码核衣壳蛋白VP1。没有观察到任何其他普通鼠病毒或密切相关的莱利斯塔德病毒和马动脉炎病毒的RNA的特异性扩增。在实验感染的小鼠中,可以在急性和持续阶段的血浆中检测到LDV。还通过PCR在被保藏在小鼠中的伯氏疟原虫的污染池中通过PCR检测了LDV,这通过样品的直接分析和通过测试来自用这些池接种的小鼠的血浆来进行。 PCR检测结果与接种小鼠血浆乳酸脱氢酶(LDH)酶检测结果完全一致。与LDH酶测定的结果相反,在PCR测定中未获得假阳性反应,阴性对照样品显示可见溶血现象。血浆样品在室温,4,-20和-80摄氏度下最多保存8天不会影响PCR结果。这些结果表明,PCR是一种有价值的技术,可以代替LDH测试作为诊断工具。

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