扩增内标及其在食源性致病菌PCR检测中的应用

虽然PCR技术不断地得到了发展和改进,但检测结果容易出现假阴性而影响检测准确性的现象一直没有得到很好的解决。现在大多数学者普遍认为,在PCR体系中加入扩增内标(即一段人工构建合成的DNA序列或者是一段致病菌的看家基因序列)能有效指示假阴性现象的出现,是PCR检测技术标准化的措施之一。本文将从PCR检测方法中假阴性出现的原因、扩增内标的构建以及扩增内标在PCR检测方面的应用三方面进行综合评述,并结合本实验室的工作基础,介绍扩增内标的简捷构建过程和应用要点,希望在不影响检测灵敏度的前提下,发挥扩增内标对假阴性的指示作用。

Internal amplification control and its applications in PCR detection of foodborne pathogens

Abstract: Despite the improvement of PCR technology, false-negative results from PCR detection remain as an unresolved issue, which may reduce the detection accuracy. To address this problem, an internal amplification control, which is a non-target DNA fragment, is sometime suggested at present to be included into a PCR detection system to be co-amplified with the target sequence so as to indicate the PCR inhibitors in the samples and false negative results in the detection. This is also necessary for the standardization of PCR detection system. The purpose of this article was to provide a review on the IAC-PCR in three aspects: the causes of the false-negative results appearing, the construction strategies of internal amplification controls and the applications of IAC-PCR. In addition, a brief introduction of related research work on IAC-PCR in our laboratory will be mentioned in this review, which might be helpful to develop an easy method for the construction of IAC, which could indicate the false-negative results without the decrease of detection sensitivity.