...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Clinical Microbiology >Evaluation of a TaqMan PCR Assay To Detect Rabies Virus RNA: Influence of Sequence Variation and Application to Quantification of Viral Loads
【24h】

Evaluation of a TaqMan PCR Assay To Detect Rabies Virus RNA: Influence of Sequence Variation and Application to Quantification of Viral Loads

机译：TaqMan PCR检测狂犬病病毒RNA的评估的评估：序列变异的影响和对病毒载量的定量应用

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Published assays that use TaqMan PCR are consistently sensitive, rapid, and readily transferable. Here we describe a TaqMan PCR-based method for the detection of rabies virus (RV) RNA in tissue samples. We show that the method has an acceptable linear range, is both sensitive and specific, and, importantly, correlates with the concentration of infectious virus. In addition, the levels of RV-specific amplification are adjustable according to the levels of an endogenous control (β-actin mRNA), allowing the calculation of comparable quantities. We tested the capacity of this assay to cope with target sequence variations. The number of sequence mismatches between gene-specific oligonucleotides and the target sequence significantly affects amplification (P < 0.001), and point mutations at the center of the probe can result in false-negative results through the prevention of probe binding and subsequent fluorescence. This study demonstrates that the genetic heterogeneity of RVs may prove a serious obstacle in the development of a diagnostic assay based on TaqMan PCR; however, the quantification of RV levels may prove to be a valuable application of this assay.
机译：使用TaqMan PCR的已发布测定始终灵敏，快速且易于转移。在这里，我们描述了一种基于TaqMan PCR的方法来检测组织样本中的狂犬病毒（RV）RNA。我们表明该方法具有可接受的线性范围，既灵敏又特异，并且重要的是与传染性病毒的浓度相关。此外，RV特异性扩增的水平可根据内源性对照（β-肌动蛋白mRNA）的水平进行调节，从而可计算相当数量。我们测试了该测定法应对靶序列变异的能力。基因特异性寡核苷酸与靶序列之间的序列错配数量显着影响扩增（ P <0.001），并且探针中心的点突变可通过防止HBsAg产生假阴性结果。探针结合和随后的荧光。这项研究表明，RVs的遗传异质性可能在开发基于TaqMan PCR的诊断检测方法中被证明是一个严重的障碍。但是，RV水平的定量可能证明是该测定的有价值的应用。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号