金黄色葡萄球菌rep-PCR指纹图谱分型技术反应条件的优化

摘要

[Objective] To optimize reaction condition of repetitive sequence based PCR ( rep - PCR) fingerprint map classification for staphylococcus aureus, and supply data to a system of S. Aureus. [Methods] Genomic DNA of S. Aureus reference strain ATCC 25923 was abstracted with bacteria DNA abstract bit and used as the template for PCR. To optimize template primer, primer concentration and annealed temperature of PCR optimized. [Results] For S. Aureus, the electro-phoresis bands of amplified products were most clear when using primer rep 1.0 pmol/μX and annealing at 45℃. [ Conclusion ] To optimum rep - PCR fingerprint map classification method of S. Aureus is obtained.%[目的]优化金黄色葡萄球菌细菌基因组重复序列PCR( rep - PCR)指纹图谱的分型方法,为构建金黄色葡萄球菌同源性追踪体系提供依据.[方法]以金黄色葡萄球菌ATCC25923基因组DNA作为模板,对参考文献提供的PCR引物,反应体系的引物浓度以及退火温度进行优化,以期得到最佳的指纹图谱.[结果]对于金黄色葡萄球菌进行指纹图谱的构建,引物rep为最佳引物,PCR反应体系引物浓度为1.0 pmol/μL,退火温度为45℃时,指纹图谱条带最清晰,明亮.[结论]优化了金黄色葡萄球菌rep-PCR分子分型技术.