不同方法检测女性生殖道病原微生物感染的比较研究

Comparative study of different detection methods for sexually transmitted infections in women

目的对比应用聚合酶链式反应(polymerase chain reaction,PCR)结合导流杂交技术检测生殖道病原微生物的DNA与应用实时荧光核酸恒温扩增技术(simultaneous amplification and testing,SAT)检测生殖道病原微生物的RNA(SAT-RNA法)的效果,探讨其临床应用价值。方法随机选取2019年12月至2020年2月期间在清华大学附属北京清华长庚医院就诊的82例疑似生殖道病原微生物感染患者作为研究对象。对82例患者均进行PCR结合导流杂交技术检测淋病奈瑟菌(neisseria gonorrhoeae,NG)、沙眼衣原体(chlamydia trachomatis,CT)、解脲脲原体(ureaplasma urealyticum,Uuu)、微小脲原体1/3/6/14(ureaplasma parvum1/3/6/14,Uup1/3/6/14)、人型支原体(M.hominis,Mh)、生殖支原体(mycoplasma genitalun,MG)和单纯疱疹病毒Ⅱ型(herpes simplex virus typeⅡ,HSV-Ⅱ);同时82例患者中,用SAT-RNA法检测70例患者的NG、76例患者的CT、34例患者的MG、29例患者的UU;以Sanger测序结果为金标准,对比用这两种方法的检测结果。结果两种方法检测NG和MG一致性的标本占100.00%;两种检测方法检测CT一致性的标本占98.68%(75/76),1例差异标本PCR结合导流杂交技术检测结果与Sanger测序结果一致;两种检测方法检测UU一致性的标本占82.76%(24/29),5例差异标本结果中,3例与PCR结合导流杂交技术检测结果相符,2例与SAT-RNA检测结果相符,且PCR结合导流杂交技术对UU进行分型,Uuu、Uup1、Uup3、Uup6、Uup14的阳性率分别为6.10%(5/82)、6.10%(5/82)、19.51%(16/82)、9.76%(8/82)、6.10%(5/82)。同时,PCR结合导流杂交技术还可以检测Mh和HSV-Ⅱ,阳性率分别为10.98%(9/82)、3.66%(3/82)。结论 PCR结合导流杂交技术检测CT、UU的特异度和准确率高于SAT-RNA法,其可有效检测出病原体的协同感染和单一感染情况,且可对UU进行分型,为判断致病性感染与正常携带、有效避免漏诊误诊提供了更精准的检测依据。

Objective To evaluate the result of using simultaneous amplification and testing(SAT) and polymerase chain reaction(PCR) flow-through hybridization to detect sexually transmitted infections and to study its clinical application value.Methods Between Dec.2019 and Feb.2020,82 patients with suspected sexually transmitted infections who were treated at Beijing Tsinghua Changgung Hospital were included as research subjects.In the 82 patients PCR-flow hybridization was used to detect the DNA of Neisseria gonorrhoeae(NG),Chlamydia trachomatis(CT),Ureaplasma urealyticum(Uuu),Ureaplasma parvum 1/3/6/14(Uup1/3/6/14),M.hominis(Mh),Mycoplasma genitalun(MG)and Herpes simplex virus type Ⅱ(HSV-Ⅱ).At the same time,of 82 patients,70 cases of NG,76 cases of CT,34 cases of MG,and 29 cases of UU "were detected by SAT-RNA.Using the Sanger sequencing results as the gold standard,the results of PCR-flow hybridization and SAT-RNA were compared.Results Two detection methods for NG and MG were consistent in 100.00%samples.Two detection methods for CT were consistent in 98.68%(75/76) samples.The PCR results of one differential sample of CT were consistent with the Sanger results.Two detection methods for UU were consistent in 82.76%(24/29)samples.In the 5 differential samples,three differential samples of UU were consistent with PCR-flow hybridization results,and two differential samples of UU were consistent with the SAT-RNA results.In addition,PCR-flow hybridization could be used to type UU,and the positive rates of Uuu,Uupl,Uup3,Uup6 and Uup14 were 6.10%(5/82),6.10%(5/82),19.51%(16/82),9.76%(8/82) and 6.10%(5/82).At the same time,PCR-flow hybridization could also be used to detect the infection of Mh and HSV-Ⅱ,and the positive rates-were 10.98%(9/82) and 3.66%(3/82),respectively.Conclusion PCR-flow hybridization has higher specificity and accuracy than SAT-RNA.PCR-flow hybridization can effectively detect the combined infection and single infection of pathogens.In addition,PCR-flow hybridization can be used to make UU typing,which provides more prevcise detection reference for distinguishing diseased infection from normal carrying,avoiding missed diagnosis and misdiagnosis.