多重PCR检测CT、MG和NG的建立及其在流产和不孕中的应用

Detection of the CT.MG and NG by the multi-PCR technique for women's sterility and abortion.

目的 :建立一种多重PCR能同时检测 3种性传播疾病病原体 :CT、MG、NG并运用于流产和不孕的研究。方法 :选择沙眼衣原体、生殖支原体、淋球菌染色体或质粒的保守序列分别设计 3对引物扩增片段长度为 473、2 81、390bp ,再分别用内切酶TaqI、EcoRI、MspI消化后得到的片段长度为 (2 92 ;181bp)、(136 ;145bp)、(2 5 0 ;140bp)加以证实。结果 :阴道拭子 :CT阳性率 2 4 7% (2 1/ 85 ) ;MG阳性率 :16 5 % (14/ 85 ) ;NG阳性率 :1.18% (1/ 85 ) ,总阳性率 :42 % (36 / 85 ) ;组织 :CT阳性率 :18.8% (18/ 96 ) ;MG阳性率 :16 .4% (16 / 96 ) ;NG阳性率 :1.0 4% (1/ 96 ) ;总阳性率 :36 .5 % (35 / 96 )。结论 :1 多重PCR辅以内切酶图谱是检测多种传播性疾病病原体的简便有效的方法 ;2 CT、MG、NG是引起女性生殖系统感染的常见病原体并导致女性不孕、流产、死胎死产的主要原因之一。

To establish a method of multi-PCR which can detect three kinds of STD'S(sexually transmited disease) pathogenic agents:CT(Chlamydia trachomatis),MG(Mycoplasma genitalium),NG(Neisseria gnorrhoeae)and be applied in investigation for woment's abortion and sterility.Methods:Three pairs primers were designed on the basis of the published nucleotide sequences of CT,MG,NG to amplify respectively 473,281,390bp DNA fragments which were enzymolysised respectively by these restriction enzyme:TaqI,EcoRI MspI to become short fragments:292 and 181bp,136 and 145bp,250 and 140bp .Results:the urethral swabs:CT positive ratio is 24.7%(21/85);MG is 16.5%(14/85),NG is 1.18%(1/85);the total positive ratio is 42%(36/85);tissue specimens:CT positive ratio is 18.8%(18/96),MG is 16.4%(16/96),NG is 1.04%,the total potivive ratio is 36.5%(35/96).Conclusion:1.The Multi-PCR assisted by restriction enzyme is a simple effective method which is able to detect three kinds of STD'S pathogenic agents:CT,MG,NG in one time;2.CT,MG,NG are gneral pathogenic agents which induce infection of female reproductive system and one of main cause which lead to women's abortion,sterility,dead fetus,stillbirth.