恒温扩增技术在男性淋球菌性尿道炎病原菌RNA检测中的应用

The application of RNA simultaneous amplification and testing in the detection of male gonococcal urethritis

目的:探讨RNA恒温扩增技术(RNA-SAT)在男性淋球菌性尿道炎病原菌RNA检测中的应用价值。方法:选取2017年6月至2018年8月温州市人民医院男性疑似淋球菌性尿道炎感染患者322例,采用RNASAT和实时荧光定量PCR技术(FQ-PCR)进行淋病奈瑟菌(NG)检测,并对结果进行统计分析。结果:NG阳性率最高为尿液RNA-SAT法35.1%(113/322),NG阳性率最低为尿液FQ-PCR法26.1%(84/322)。尿液RNA-SAT法、尿液FQ-PCR法与分离培养法的阳性率比较,差异有统计学意义(χ2=6.312,P<0.05);而拭子RNA-SAT法、拭子FQ-PCR法与分离培养法的阳性率比较,差异无统计学意义(χ2=1.039,P>0.05)。以分离培养法作为参考,尿液RNA-SAT法检测NG的灵敏度最高为97.9%(93/95),尿液FQ-PCR法的灵敏度最低为81.1%(77/95);根据ROC曲线分析,拭子RNA-SAT法和尿液RNA-SAT法的曲线下面积(AUC)中等偏上,具有较高的诊断价值(AUC>0.9);对其中用药7 d后临床症状、体征消失且尿常规白细胞阴性的81例患者采样复查,尿液RNA-SAT法、尿液FQ-PCR法以及分离培养法的阴转率差异无统计学意义(χ2=2.531,P>0.05);拭子RNA-SAT法、拭子FQPCR法以及分离培养法的阴转率差异也无统计学意义(χ2=2.933,P>0.05)。结论:RNA-SAT法和FQ-PCR法均可作为判断NG病原菌感染的指标,且RNA-SAT法可采用尿液作为待检样本,具有取样方便、耗时短、污染小及反应稳定等优点,可用于男性淋球菌尿道炎NG检测与疗效预后评估。

Objective:To evaluate the application value of RNA simultaneous amplification and testing technology in the detection of male gonococcal urethritis.Methods:From June 2017 to August 2018,a total of 322 male patients with suspected gonococcal urethritis infection in Wenzhou People’s Hospital were detected by RNA simultaneous amplification and testing(RNA-SAT)and real-time fluorescence quantitative PCR(FQPCR),then the results were statistically analyzed.Results:The highest NG positive rate was 35.1%(113/322)by urine RNA-SAT method,and the lowest NG positive rate was 26.1%(84/322)by urine FQ-PCR method.The difference of positive rates in urine RNA-SAT,urine FQ-PCR and isolation culture was statistically significant(χ2=6.312,P<0.05).The difference of positive rates in swab RNA-SAT,swab FQ-PCR and isolation culture was not statistically significant(χ2=1.039,P>0.05).The highest sensitivity of urine RNA-SAT was 97.9%(93/95),and that of urine FQ-PCR method was 81.1%(77/95).According to ROC curve,swab RNA-SAT and urine RNASAT method have a relatively high diagnostic value(AUC>0.9).After 7 days of treatment,clinical symptoms in 81 patients disappeared and negative leukocyte were re-examined.The difference in negative rates in urine RNASAT,urine FQ-PCR and isolation culture was not statistically significant(χ2=2.531,P>0.05).The difference in negative rates of swab RNA-SAT,swab FQ-PCR and isolation culture was also not statistically significant(χ2=2.933,P>0.05).Conclusion:Both RNA-SAT and FQ-PCR can be used to diagnose NG infection;however,RNA-SAT has the advantages of convenient sampling,short time,little pollution and accurate results.So it can be used to detect NG in male gonococcal urethritis and to evaluate the effect and prognosis.