定量PCR联合基因芯片检测腹水细菌16SrRNA诊断自发性细菌性腹膜炎

目的 探讨定量PCR联合基因芯片检测腹水细菌16SrRNA基因诊断自发性细菌性腹膜炎(SBP)的意义.方法 采用实时荧光定量PCR联合基因芯片检测76例临床疑似SBP肝病患者和6例对照的非感染性腹腔积液肝病患者腹水细菌16SrRNA基因,与腹水细菌培养同时比较.结果76份疑似SBP患者腹水标本中,定量PCR联合基因芯片检测阳性17份,阳性率为22.4%,其中革兰氏阳性菌8份、革兰氏阴性菌9份;腹水细菌培养阳性6份,阳性率为7.9%,均为革兰氏阴性菌,两种方法比较,x2=18.05,P＜0.01,差异有统计学意义.两种方法检测腹水细菌阳性的6份标本,菌株鉴定结果相一致.对照病例细菌检测结果呈阴性.结论 定量PCR联合基因芯片检测腹水细菌16SrRNA基因,较腹水细菌培养的敏感性和特异性高;不仅能作出快速诊断,还能确定SBP所感染的病原菌,具有实际应用价值.

Abstract：

Objective To evaluate the significance of determining ascitic bacterial16S rRNA by quantitative PCR combined with microarray (PCR-microarray) in the diagnosis of spontaneous bacterial peritonitis (SBP). Methods Ascitic bacterial 16SrRNA was determined by real time fluorescent quantitative PCR-microarray in 76 cases of suspected SBP and 6 cases of non-infectious ascites with chronic liver diseases.The results were compared with ascitic bacterial culture simultaneously. Results Of 76 ascitic samples, 17were detected bacteria positive by PCR-microarray, including 8 Grams positive(G+) and 9 Grams negative (G-), which was higher than that by bacterial culture which had only 6 ascitic samples detected positive (all G-); the positive rates were 22.4% vs 7.9%, respectively (P ＜ 0.01). The bacterial strains detected by both methods in 6 cases had a consistency with each other. No bacteria were detected in another 6 cases of noninfectious ascites with chronic liver diseases. Conclusions Determination of ascitic bacteria 16S rRNA by PCR-microarray has a higher specificity and sensitivity in the diagnosis of SBP as compared with the bacteria culture. Application of this novel method can not only accelerate SBP diagnosis but also stratify the different pathogens.

Determination of ascitic bacterial 16S rRNA by quantitative PCR-microarray in the diagnosis of spontaneous bacterial peritonitis

Objective To evaluate the significance of determining ascitic bacterial16S rRNA by quantitative PCR combined with microarray (PCR-microarray) in the diagnosis of spontaneous bacterial peritonitis (SBP). Methods Ascitic bacterial 16SrRNA was determined by real time fluorescent quantitative PCR-microarray in 76 cases of suspected SBP and 6 cases of non-infectious ascites with chronic liver diseases.The results were compared with ascitic bacterial culture simultaneously. Results Of 76 ascitic samples, 17were detected bacteria positive by PCR-microarray, including 8 Grams positive(G+) and 9 Grams negative (G-), which was higher than that by bacterial culture which had only 6 ascitic samples detected positive (all G-); the positive rates were 22.4% vs 7.9%, respectively (P ＜ 0.01). The bacterial strains detected by both methods in 6 cases had a consistency with each other. No bacteria were detected in another 6 cases of noninfectious ascites with chronic liver diseases. Conclusions Determination of ascitic bacteria 16S rRNA by PCR-microarray has a higher specificity and sensitivity in the diagnosis of SBP as compared with the bacteria culture. Application of this novel method can not only accelerate SBP diagnosis but also stratify the different pathogens.