葡萄球菌染色体mec盒多位点复合PCR分型测定

Multilocus Complex PCR Typing of Staphylococcus Chromosome Cassette mec

目的研究用多位点复合PCR分型法高效、快速、准确地测定金黄色葡萄球菌SCCmec型别。方法采用正交设计和平行对比分析,对影响测定结果的退火温度、延伸时间,MgCL2、dNTP、Taq DNA聚合酶、引物的用量进行了测试。结果获得多位点PCR测定的最佳退火温度为55℃、延伸时间为1 min,以及MgCl2、dNTP、TaqDNA聚合酶和各位点引物的最佳用量。结论多位点PCR测定是一种高效、快速、实用的SCCmec分型方法,可用于大规模的MRSA分子流行病学研究。

OBJECTIVE For high effectively, quickly and exactly identifying the SCCmec types of Staphylococcus aureus with a multilocus complex PCR typing. METHODS Using the orthogonal design and parallel control design to test the effect of annealing temperature, extensioa time and the concentration of MgC12, dNTP, Taq DNA polymerase and primers on the result. RESULTS Itrs turned out that the perfect annealing temperature was 55 ℃, extension time was 1 minute, and also got the perfect concentration of MgC12, dNTP, Taq DNA polymerase and primers. CONCLUSIONS The multilocus complex PCR is a practical method for effectively and quickly identifying the SCCmec types of MRSA and can be used in molecular epidemiologic study with a large scale.