聚合酶链反应法快速诊断肝硬化患者腹水细菌感染临床应用研究

Clinical application of PCR in the quick diagnosis of bacterial infection of ascites in patients with cirrhosis

目的应用聚合酶链反应（PCR）检测细菌16SrRNA基因,探讨其在辅助诊断肝硬化自发性腹膜炎（SBP）中的临床应用价值。方法选取2014年12月-2015年10月医院感染病科77例肝硬化失代偿期合并腹水患者进行研究,以细菌16SrRNA基因为靶序列,在保守区和特异区分别选取特异性序列合成引物,建立快速诊断方法,检测临床肝硬化失代偿期患者腹水中细菌DNA,并将其结果与传统方法进行比较,数据采用SPSS16.0统计软件进行处理。结果 77份临床样本中PCR法16SrRNA通用引物阳性扩增50例,阳性率64.9%,特异引物阳性扩增15例,阳性率19.5%;以腹水细菌培养阳性或（和）腹水PMN≥250×106/L或（和）腹膜刺激征阳性且正规抗感染治疗有效者作为诊断SBP金标准,此时PCR法灵敏度为100.0%,特异度为54.0%,阳性预测值为54.0%,阴性预测值为100.0%,二者差异有统计学意义（χ~2=22.453,P〈0.01）。结论与传统方法相比,PCR法是一种更敏感且简便快速的方法,在肝硬化失代偿期患者腹水细菌检测中具有一定的临床应用价值。

OBJECTIVE To evaluate the clinical application value of polymerase chain reaction（PCR）in the diagnosis of spontaneous bacterial peritonitis（SBP）in patients with liver cirrhosis through the detection of 16 SrRNA.METHODS A total of 77 patients with liver cirrhosis in decompensated period combined ascites in our hospital from Dec.2014 to Oct.2015 were selected.Using bacterial 16 SrRNA gene as the target sequence,specific sequences were selected to establish a rapid diagnostic method in the universal and specific region.The bacterial DNA in ascites of the patients with liver cirrhosis was detected by this method,and the results were compared with the conventional methods.SPSS 16.0was used for data analysis.RESULTS From the 77 submitted specimens,50cases（64.9%）were positively amplified by 16 SrRNA universal sequence and 15cases（19.5%）by specific sequence through PCR method.Taking ascites culture positive,PMN≥250×106/L or（and）peritoneal irritation sign positive and anti-infection treatment effective as SBP diagnostic gold standard,in which the sensitivity of PCR method was 100.0%,the specificity 54.0%,the positive predictive value 54.0%,the negative predictive value 100.0%,and the difference between the two groups was significant（χ~2=22.453,P〈0.01）.CONCLUSIONCompared with the traditional methods,the PCR assay is a more sensitive and simple method which is a certain value of the detection of ascites in patients with liver cirrhosis.