PCR的优化与发展  

作　　者：刘姗姗[1] 岳素文 江洪 王成彬[3] 

机构地区：[1]温州医科大学检验医学院,生命科学学院,浙江温州325035 [2]北京泰格瑞分子检验有限公司,北京102209 [3]中国人民解放军总医院临床检验科,北京100853

出　　处：《临床检验杂志（电子版）》2014年第2期610-617,共8页Clinical Laboratory Journal(Electronic Edition)

摘　　要：实时荧光定量PCR亦成为RT-QPCR的出现,确定了PCR(聚合酶链)反应的技术实现由了定性检测到兼定量检测靶标核苷酸序列的华丽变身,是对PCR质变式的优化,但是该技术在现实应用中依然存在着大量的难以克服的障碍:分析溶解曲线的过程耗时很长、标记任意荧光探针的费用昂贵、非特异性的扩增干扰等,长时间来都阻碍着RT-QPCR的技术应用。迄今,PCR中隐含的这些难题的具体机理仍未被彻底弄明白,所以无法从彻底被消除,因此多种优化PCR的技巧需要被采纳以推动应用方面和在其有关的领域的研究发展。因此,PCR技术中有关优化方案的研究发展迅速,这不仅仅能够提高PCR方法的扩增效率,还能有针对性地抑制非特异性扩增,PCR定量分析、检测技术的适用范围被无限推广,而且对于分子生物学的核心技术的发展而言也有利于其顺利地发展。本文就目前对PCR优化与发展的近期进展进行了论述和展望。[Objective] The appearance of Real-time fluorescent quantitative PCR(RT-QPCR) marked the polymerase chain reaction(PCR) technology magnificently turning from the qualitative to adding the quantitative detection of targeted specific genes, which is a great optimization of conventional PCR. However, there still exists lots of insurmountable problems in the practical applications of it, such as the nonspecific amplification phenomenons, fluorescent probes are too much expensive now, the dissolution curves analysis is time-consuming, which all block RT-QPCR's applications for a long time. At present, the mechanisms are still not clear for the occurrence of these problems existing in PCR, hence we cannot solve these problems abosolutly. Therefore, we need to adopt a variety of solutions for optimizing PCR to promote the applications of this technology and the progress of these research fields. As a result, the studies of optimization schemes for the PCR technology has obtained great development, which has not only improved the amplification efficiency of PCR, but also can inhibit nonspecific amplification effectively and promote quantitative PCR detection. Last but not the least, it also palyed an important role in the applicable scope as well as improved the research in the fields of molecular biology techniques. In this paper, the current research progress of PCR optimization schemes is discussed and prospected.

关 键 词：PCR ARMS 扩增 引物二聚体 

分 类 号：R440[医药卫生—诊断学]