细菌革兰双重16S rRNA基因荧光定量PCR快速诊断妇产科菌血症

摘要

目的:用已知细菌革兰阴性、阳性菌双重16S rRNA基因荧光定量PCR方法(q-PCR)应用于妇产科患者菌血症的诊断,探讨该方法检测妇产专科患者菌血症的临床应用价值.方法:2013年1月～2014年12月对100例疑为全身感染处于菌血症状态的住院分娩孕产妇进行常规血液培养,同时用细菌革兰阴性、阳性菌双重16SrRNA基因q-PCR方法检测,分析2种方法诊断菌血症的阳性率、敏感性和特异性.结果:通过q-PCR方法检测菌血症阳性率44％,血液培养阳性率16％,两者差异有统计学意义(P＜0.01);菌血症临床诊断阳性率为37％,与q-PCR法阳性率存在显著性差异(P＜0.01).以血液培养阳性和(或)临床诊断菌血症的标准作为对照,q-PCR方法诊断敏感性为89.2％,特异性82.5％.结论:细菌革兰阴性、阳性菌双重16S rRNA基因q-PCR方法检测妇产科菌血症的阳性率高于血液培养,可快速为孕产妇患者感染提供早期、敏感的病原学诊断依据.%Objective:To develope a fluorescence quantitative polymerase chain reaction (FQ-PCR)method based on 16S rRNA genes for the diagnosis of infections in gynecological and obstetric patient in order to improve the efficiency and accuracy of bacterial detection.Methods:One hundred patients suspected with bacterimia were cultured and bacterial 16S rRNA gene were detected synchronously by extraction of DNA,primer and probe design,PCR amplification and fluorescent quantitative detection of amplified products.The positive rate,sensitivity and specificity of these two methods were compared.Results:The positive rate was 44％ by FQ-PCR,which was significantly higher than 16％ by blood culture (P＜0.01).Taking the criteria of positive blood culture and/or clinical diagnosis of bacterimia as control,the sensitivity was 89.2％ and the specificity was 82.5％ for FQ-PCR.Conclusion:The positive rate of FQ-PCR was significantly higher than that of blood culture and could be an early and sensitive method for pathogenic diagnosis of bacterimia in gynecological and obstetric patients.