Standardization and quality control of PCR analyses
Standardization and quality control of PCR analyses
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DOI：
10.1515/cclm.2000.014
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发表时间：
2000-02-01
影响因子：
6.8
通讯作者：
Burkardt, HJ
中科院分区：
文献类型：
--
作者：
Burkardt, HJ
在聚合酶链式反应(PCR)检测进入感染性病原体诊断领域的一开始，该技术因其经常因污染而产生假阳性结果的倾向而阻碍了该技术在常规诊断中的引入。现在，通过引入尿嘧啶-N-糖基酶(UNG)抗污染技术，这个问题得到了广泛的解决。然而，仍然必须注意避免产生假阳性结果的其他来源。它们还可能源于人为错误和/或不充分的聚合酶链式反应扩增和检测方案。一种特殊的情况是，聚合酶链式反应还会放大死亡生物的DNA，使诊断正确的结果成为阳性，但在临床上却是假阳性。在聚合酶链式反应中，就像在任何其他诊断测试中一样，也存在产生假阴性结果的风险。在这种情况下，除了人为错误、低靶点或糟糕的扩增和检测方案外，最有可能的来源是患者样本中干扰物质造成的抑制。本文讨论了认识和克服这一问题的策略。最后，对扩增技术在感染性病原体诊断中的一些质量控制研究结果进行了综述。
In the very beginning of polymerase chain reaction (PCR) tests entering the field of diagnosis of infectious agents, the introduction of this technology into routine diagnosis was hampered by its frequent tendency to create false-positive results because of contamination. This problem is now widely solved by the introduction of the uracil-N-glycosylase (UNG) anticontamination technology. However, care must still be taken to avoid other sources of producing false positive results. They might additionally derive from human error and/or insufficient PCR amplification and detection protocols. A special case lies in the fact that PCR also amplifies DNA from dead organisms rendering a result diagnostically correct as positive, but clinically as false-positive. In PCR, as in any other diagnostic test, the risk of creating a false-negative result also exists. In such a case, the most probable source besides human error, low target or poor amplification and detection protocols is an inhibition caused by interfering substances in a patient's sample. Strategies to recognize and overcome this issue are discussed in this article. Finally a few results from quality control studies on amplification technologies in the diagnosis of infectious agents are reviewed.