PCR和传统培养法检测腹膜透析相关性腹膜炎致病菌的对比分析  

Comparison of polymerase chain reaction and bacteria culture in peritoneal dialysis associated peritonitis

作　　者：王婵媛[1] 吴永贵[1] 齐向明[1] 王娟[1] 戴宏[1] 卢文[1] 

机构地区：[1]安徽医科大学第一附属医院肾内科,合肥230022

出　　处：《中华肾脏病杂志》2015年第12期898-904,共7页Chinese Journal of Nephrology

摘　　要：目的 探讨PCR检测法在腹膜透析相关性腹膜炎（PDAP）细菌检测中的临床应用价值.方法 收集2014年1月至2014年12月安徽医科大学第一附属医院PDAP患者腹膜透出液标本40份,分别行传统细菌培养与PCR法检测致病菌.根据细菌16S rRNA基因设计细菌通用引物,并参考文献设计17种细菌的特异引物.提取腹透液DNA后行实时荧光定量PCR （qPCR）扩增.设立标准菌株DNA提取液为阳性对照,无菌双蒸水作为阴性对照.结果 （1）传统细菌培养法结果阳性26例（26/40）,革兰阳性球菌18例（18/26）,主要菌种表皮葡萄球菌5例、溶血葡萄球菌4例、大肠杆菌4例、金黄色葡萄球菌3例.（2）PCR检测细菌结果阳性为33例（33/40）,主要菌种与普通培养结果无异;阳性标本中有2例未测出具体菌种.（3）以细菌培养作为金标准,PCR检测技术对PDAP致病菌检测的敏感度为96.15％,特异度为42.86％,且检出阳性率明显高于普通培养法,组间比较差异有统计学意义（P＜0.05）.（4）PCR法检测致病菌4～6h内出结果,普通培养平均报告时间为（77.88±15.53）h,明显长于PCR法.结论 与传统细菌培养法比较,PCR检测法具有敏感、简便、快速的优点,在PDAP患者腹透液细菌的检测中具有一定的临床价值.Objective To evaluate clinical application value of polymerase chain reaction （PCR） detection for bacteria in peritoneal dialysis associated peritonitis （PDAP）.Methods Peritoneal dialysis fluid specimens were collected from January 2014 to December 2014 in The First Affiliated Hospital of Anhui Medical University.Conventional bacterial culture and PCR detection were used respectively.According to the bacterial 16S rRNA gene, universal primers were devised and designed, based on reference, the specific primers of 17 kinds of experimental bacteria.Real-time fluorescent PCR （Real-time PCR, qPCR） amplification was implemented.The establishment of standard strain DNA extract was used as positive control;sterile double distilled water was used as negative control.Results （1） The traditional bacterial culture results showed that positive proportion was 26/40 in specimen of 40 cases, gram-positive strains accounting for 18/26.Main species were epidermis staphylococcus （5/26）, hemolysis staphylococcus （4/26）, escherichia coli （4/26）, and streptococcus viridans （3/26）.（2） The PCR detection results showed that total positive rate was 33/40 in 40 patients specimens, among which 2 cases of positive samples ended up with no specific strains being detected;the main bacteria strains in PCR were not different from ordinary culture results.（3） With bacterial culture as the gold standard, the detection sensitivity of PCR technology for PDAP pathogenic bacteria was 96.15% and specificity was 42.86%;the detection positive rate was significantly higher than ordinary culture method.（4） PCR technology for detecting pathogenic bacteria could produce results within 4-6 hours, while reported positive results in the traditional bacterial culture would take （77.88±15.53） hours, which was significantly longer than PCR.Conclusion Compared with traditional bacteria culture method, PCR method is more sensitive, simple, and quick.Bacteria detection using PCR technique is of clinic applied value in

关 键 词：腹膜炎 腹膜透析 致病菌 聚合酶链反应 普通培养 

分 类 号：R446.5[医药卫生—诊断学]