摘要
目的探索快速可靠的新生儿败血症和化脓性脑膜炎诊断的新方法及其临床运用.方法用自行设计的细菌16S rRNA基因高度保守区的引物和探针,对19种标准菌株、3种病毒株、新型隐球菌及白色念珠菌、人基因组DNA,以及疑似败血症的临床血标本、脑脊液共195份进行荧光定量检测,并同时进行血或脑脊液培养的对照.结果细菌荧光定量PCR具有较好的敏感性和特异性,最低能检测到10个拷贝的16S rRNA基因,即相当于1～2个细菌,与人基因组DNA、真菌及病毒等无交叉阳性反应;荧光定量PCR的细菌DNA检出率为12.8%(25/195),明显高于血培养的阳性率7.1%(15/195,P＜0.01).结论应用通用引物和探针的荧光定量PCR扩增诊断败血症的方法检测快速、敏感性和特异性高,具有较大的推广及应用价值.
Objective To explore a rapid diagnostic method in neonatal sepsis and bacterial meningitis. Methods The primers and TaqMan probes were designed and synthesized based on the sequences of bacterial 16S rRNA gene. Nineteen bacterial strains, 3 different viruses, fungus and human genomic DNA were tested by FQ-PCR assay. Blood specimens and CSF from 195 cases of suspected septicemia were detected with both TaqMan PCR assay and blood or/and CSF culture. Results The FQ-PCR showed very high sensitivity and specificity and was able to detect at least 10 copies of 16S rRNA gene equivalent to 1 - 2 copies bacterium. No cross-reaction was found with human genomic DNA, other fungus and viruses. Among the 195 cases, the positive rate by FQ-PCR was 12.8%（25 cases） and 7. 1% （15 cases） by blood culture （P〈0.01）. Conclusions Real-time PCR for sepsis diagnosis was established and characterized by its rapidity, sensitivity and specificity which might have great value in clinical practice.
出处
《中华围产医学杂志》
CAS
2005年第4期242-245,共4页
Chinese Journal of Perinatal Medicine
