摘要
目的建立基于16SrRNA基因的聚合酶链反应(PCR)细菌学检测方法,以指导临床快速诊断新生儿败血症。方法以细菌16SrRNA基因为靶序列,利用计算机软件设计合成所有细菌保守区共有的一对通用引物,通过PCR扩增已知10株实验室保存菌株、人类基因组DNA、巨细胞病毒、白色假丝酵母菌和空白对照,并对其灵敏度和特异性作一评价。结果对所测已知10株实验室保存菌株均获得920bp扩增产物,其与人类基因组DNA、巨细胞病毒和白色假丝酵母菌无交叉阳性反应;PCR阳性率为26.8%,血培养阳性率为11.3%,两者比较有差异有统计学意义(P<0.05),PCR灵敏度高于血培养。结论与血培养比较,PCR具有检测速度快、特异性及敏感度高等特点。
OBJECTIVE To establish PCR method for the bacteriology detection based on 16S rRNA gene,in order to explore a method for the rapid diagnosis of neonatal septicemia.METHODS We designed and synthesized a pair of universal primers by computer software as a target sequence of all bacterial region.16S rRNA fragments from 10 known strains of laboratory strains were amplified with PCR by using human genomic DNA,cytomegalovirus,Candida albicans and blank-control as the controls,its sensitivity and specificity was evaluated.RESULTS Totally 920bp amplified products were acquired from the 10 reserved laboratory strains,without cross-positive reactions with human genomic DNA,Cytomegalovirus,and C.albicans;the positive rate of PCR was 26.8%,the positive rate of blood culture was 11.3%,the difference was statistically significant（P0.05）,the sensitivity of PCR is higher than that of blood culture.CONCLUSION As compared with blood culture,the detection with PCR is rapid with high specificity and sensitivity.
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2012年第21期4696-4697,4700,共3页
Chinese Journal of Nosocomiology
