摘要
目的 建立一种利用MCSP的PCR扩增产物快速检测和鉴别细菌的方法。方法 分析 3种致病菌MCSP序列 ,针对不同细菌的MCSP的冷休克域的基因序列 ,设计了一对通用PCR引物 ,直接以细菌体为模板 ,相同条件下同时扩增多种细菌的MCSP基因序列。结果 对TouchdownPCR条件进行优化后 ,同时扩增出 3种细菌MCSP基因序列 ,经过电泳和基因测序 ,确认PCR扩增产物即为扩增靶序列。结论 具有高度同源性不同种细菌的MCSP基因片段之间存在足够的碱基差异 ,将细菌鉴别至种 ,甚至鉴别至属。与 1 6srRNA相比较更具有实用价值。与基因芯片技术相结合 。
Objective To develop a diagnostic technique to detect and identify bacteria quickly.Methods Using two universal PCR primer oligomers from conserved regions of MCSP DNA sequence homologues to amplify a 200 base pair DNA sequence of bacteria,including escherichia,staphyloccus and pseudomonas.Results Using the two primers,we amplified partial MCSP gene sequences directly.Conclusion Although MCSPs DNA sequence shows considerable homology,there is sufficient nucleotide variation to allow the unique allocation of each amplified product to its parental bacterium.Compared to 16s rRNA,targeting MCSP nucleic acid sequences has more potential for the discrimination of live bacteria from those that non viable and dead.This assay has a bright future development collaborating with gene chip in the field of clinical detection.
出处
《重庆医学》
CAS
CSCD
2004年第8期1126-1128,共3页
Chongqing medicine
