Evaluation of minor groove binding probe and Taqman probe PCR assays:: Influence of mismatches and template complexity on quantification
Evaluation of minor groove binding probe and Taqman probe PCR assays:: Influence of mismatches and template complexity on quantification
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DOI：
10.1016/j.mcp.2006.03.003
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发表时间：
2006-10-01
影响因子：
3.3
通讯作者：
Karlsson, Hakan
中科院分区：
文献类型：
--
作者：
Yao, Yuanrong;Nellaker, Christoffer;Karlsson, Hakan
使用3 '-小沟结合物（MGB）或Taqman探针的实时PCR测定广泛用于临床病毒学测试和突变/多态性检测。我们比较了3 '-MGB探针与常规Taqman探针的线性、灵敏度、特异性和动态范围。使用含有不同错配的质粒或使用人基因组DNA作为模板比较两种测定法的性能。观察到MGB和Taclman探针测定的线性和灵敏度相当。使用标准条件，没有一种测定是序列特异性的。在Taqman探针测定中，多达五个错配产生可检测的信号。Taqman和MGB探针测定的性能受模板复杂性的影响，然而，后者的影响程度较小。总之，这些结果突出了MGB探针在错配识别方面优于Taqman探针的优点，但表明反应条件的优化和特异性的验证对于MGB探针也是必要的。(c)2006爱思唯尔有限公司保留所有权利。
Real-time PCR assays using 3'-minor groove binder (MGB) or Taqman probes are widely used for clinical virological testing and mutation/polymorphism detection. We compared a 3'-MGB probe to a conventional Taqman probe for linearity, sensitivity, specificity and dynamic range. The performance of the two assays was compared using plasmids containing different mismatches or using human genomic DNA as a template. Comparable linearity and sensitivity were observed for the MGB and the Taclman probe assays. Using standard conditions, none of the assays were sequence- specific. Up to five mismatches generated a detectable signal in the Taqman probe assay. The performance or the Taqman as well as the MGB probe assay was influenced by the complexity of the template, the latter, however, to a lesser degree. Overall, these results highlight the advantages of the MGB probe over the Taqman probe regarding mismatch discrimination, but suggest that optimization of reaction conditions and verification of the specificity are necessary also for MGB probes. (c) 2006 Elsevier Ltd. All rights reserved.