关节液16S rRNA实时逆转录PCR在假体周围感染诊断中的应用

摘要

Objective To investigate the efficiency of 16S rRNA Real-time reverse transcription PCR technique in the diagnosis of periprosthetic joint infection,and compare its sensitivity and specificity with conventional culture.Methods There were 43 revision cases from July 2013 to December 2015.Synovial fluid collected by puncture preoperatively,tissues from five different parts around the prosthesis collected intra-operatively were cultured by blood plate and BacT/Alert FN respectively.The 16S rRNA in interface membrane was detected by real-time reverse transcription PCR as a marker to diagnose PJI.At the same time,the synovial fluid was routinely bacterial cultured.We compared the sensitivity and specificity of two methods.Results There are 22 THAs and 21 TKAs respectively in 43 cases,23 cases diagnosed prosthetic joint infection and 20 cases diagnosed non prosthetic joint infection.The sensitivity of 16S rRNA Real-time reverse transcription PCR is higher than the conventional bacterial culture (78.2％ vs.47.8％).There was no difference in the specificity and PPV and NPV.For PCR in prosthetic joint infection group,Staphylococcus epidermidis in 5 cases,Staphylococcus aureus in 3 cases,streptococcus in 4 cases,E.coli in 2 cases,Staphylococcus lugdunensis,Pseudomonas aeruginosa,Staphylococcus haemolyticus and Mycoplasma in 1 case respectively.For culture in prosthetic joint infection group,Staphylococcus epidermidis in 5 cases,Staphylococcus aureus in 2 cases,Staphylococcus lugdunensis,Pseudomonas aeruginosa,Staphylococcus haemolyticus and E.coli in 1 case respectively.For non prosthetic joint infection group,PCR and culture are all negative.Conclusion The sensitivity of 16S rRNA Real-time reverse transcription PCR is higher than the conventional bacterial culture.%目的 探讨关节液16S rRNA实时逆转录PCR技术在假体周围感染中的诊断效率,并比较其与常规培养的敏感性和特异性.方法 2013年7月至2015年12月接受人工关节翻修手术的43例患者,术前行关节液穿刺,术中采集5个高度可疑的假体周围组织,将术前关节液、术中假体周围组织分别进行血平板和BacT/Alert FN瓶培养,并采用16SrRNA实时逆转录PCR技术检测关节液中的细菌,分析其对假体周围感染的诊断效率.比较关节液16S rRNA实时逆转录PCR与常规培养在细菌检出率的差异.结果 43例患者中感染组23例,非感染组20例.全髋关节22例,全膝关节21例.关节液16S rRNA实时逆转录PCR技术的微生物检出敏感性(78.2％)高于常规细菌培养(47.8％),两者的特异性、阳性预测值及阴性预测值相当.确诊为假体周围感染的23例中PCR阳性结果为表皮葡萄球菌5例,金黄色葡萄球菌3例,链球菌4例,大肠杆菌2例,路邓葡萄球菌、铜绿假单胞菌、溶血性葡萄球菌及人型支原体各1例;培养阳性结果为表皮葡萄球菌5例,金黄色葡萄球菌2例,路邓葡萄球菌、铜绿假单胞菌、溶血性葡萄球菌及大肠杆菌各1例.20例非感染组中PCR和常规培养均阴性.结论 关节液16S rRNA实时逆转录PCR可以提高假体周围感染病原微生物检出率,对于关节置换术后假体周围感染的诊断有一定的参考价值.