标题：实时荧光PCR和普通PCR方法检测病原微生物的灵敏度比较

      作者：钟 岸，蔡 蓁，王 毅
    （深圳市孙逸仙心血管医院检验科，广东 深圳 518020）

      卷次： 2013年24卷13期

      【摘要】 目的 通过比较两种 PCR检测的灵敏度，选择不同条件下经济准确的方法检测病原微生物。
方法 制备标准样品用以两种PCR扩增条件的确定，通过目的基因片段设计引物，扩增菌体裂解液上清。PCR
产物分别通过扩增曲线和琼脂糖电泳检测目的条带的存在，以最低检测的条带浓度作为两种不同PCR检测的最
高灵敏度，通过比较，选择在不同的限制条件下较为方便经济的方法检测病原微生物。结果 实时PCR中裂解
菌液上清经过10-8倍稀释后仍能观测到扩增曲线的存在。而普通PCR产物经过琼脂糖电泳染色后观测只能观测
到10-5浓度的条带，灵敏度远不及RT-PCR。结论 荧光PCR检测灵敏度较高，普通PCR琼脂糖电泳方法只能达
到RT-PCR检测灵敏度的一半但比较经济。

      【关键词】 实时PCR；病原微生物；灵敏度

      【中图分类号】 R446.5 【文献标识码】 A 【文章编号】 1003—6350（2013）13—1956—03


Comparison of the sensitivity of RT-PCR and general PCR in detecting pathogenic microorganisms.
ZHONG
An, CAI Zhen, WANG Yi. Department of Clinical Laboratory, Shenzhen Sun Yat-sen Cardiovascular Hospital, Shenzhen
518020, Guangdong, CHINA

【Abstract】 Objective To compare the detecting threshold of two different PCR methods, and to select the
accurate and economical way to detect the pathogenic microorganisms. Methods Standard samples were applied to
establish the optimal condition for the two kinds of PCR methods. Primer design was based on the targeting sequence
of the pathogenic microorganism. The products of the PCR were detected by amplification plot or agarose-gel electro-
phoresis. The highest detecting sensitivity was set by the lowest concentration which is able to be detected by PCR. By
comparison, the optimal method was chosen for the pathogenic microorganism detection. Results Supernatant of
bacteria lysis solution in real-time PCR after 10-8 dilution can still be observed with amplification curve. The general
PCR products after agarose gel electrophoresis staining can be observed only with the concentration of 10-5. The sensi-
tivity of general PCR was significantly lower than that of RT-PCR. Conclusion RT-PCR has higher detecting sensi-
tivity, but general PCR is more economical.

      【Key words】 RT-PCR; Pathogenic microorganisms; Sensitivity