parC检测在解脲支原体基因分型中的临床应用

Clinical application of genotyping detection of human Mycoplasma urealyticum based on parC gene sequences

目的根据parC基因序列差异建立一种解脲支原体基因分型方法，以实现其两个生物型Uu（Ureaplasma urealyticum）和up（Ureaplasma parvum）的快速鉴定，便于临床常规应用。方法依据解脲支原体14个血清型标准株parC基因序列，设计可区分观和印的特异性引物与和探针；通过检测serovarl和serovat4标准株、50份解脲支原体临床分离株（Uu12株，Up38株），7种阴道常见分离菌以鉴定该方法的特异性；留取70例性病门诊非淋球菌性泌尿生殖道炎症患者和71例妇科正常体检者的标本，分别进行解脲支原体培养和基因分型鉴定，以比较两种方法的敏感性，并应用统计学分析Uu、Up在性病和正常人群中分布的差异。结果应用parC基因分型法成功将serovarl标准株、serovat4标准株、Uu和Up临床分离株鉴别，Uu、Up两生物群之间以及7种阴道常见菌均未出现非特异性扩增；基因分型方法的敏感性显著高于培养法（P〈O．05）；在70例性病门诊患者中，Uu和Up的检出率分别为8．57％和61．40％，两者混合感染为24．30％；在7l例妇科体检人群中魄和唧检出率分别为7．04％和67．60％，两者混合感染为8．45％，性病门诊病人中解脲支原体感染率显著高于体检人群（P〈O．05），Uu和Up单纯感染在两群体中的分布差异无统计学意义（P〉O．05），而混合感染在性病门诊病人中显著增加（P〈O．05）。结论解脲支原体在性病患者中的感染率显著高于健康体检者，且混合感染在性病患者中明显增加，而Uu和Up单纯感染在两人群中分布差异无统计学意义，因此解脲支原体的致病可能与其不同基因型的混合感染有关。

Objective To establish a novel PCR method that can differentiate the two biotype of Mycoplasma urealytium rapidly based on the disparities of parC gene sequences for clinical routine examination. Methods Design two pairs of specific primers and probes for the targeted gene according to the differences of the parC gene sequences from 14 standard serotypes. The specificity of this amplification was verified by detecting two Mycoplasma urealyticum standard strains including serovarl and serovar4, 50 clinical isolated strains （ 12 are Uu strains and 38 are Up strains） and 7 common baeterias from vagina. Collected 70 swab specimens from patients of Sexually Transmit Department with urogenital inflammation symptom and 71 swab specimens from the gynecology health examination population. All those specimens were detected by culture and our method respectively. The sensitivity of this method was evaluated by comparing with the culture. Differences of the infection rate and distribution of biotypes between different populations were analyzed using statistical software. Results Standard strains of Mycoplasma urealyticum and clinical isolates can be typed into two species by the PCR method without nonspeeifie amplification. The sensitivity of PCR method is much higher than that of culture（ P〈0.05 ）. The infection rates of Uu, Up and the mixing were 8.57% , 61.40% , 24.30% respectively for the patients with vaginitis. However, it was 8.80% , 67.60% , 8.45% respectively for the gynecology health examination population. There is a significant difference of the total infection rate between the two oooulation（ P〈0.05 ）. It showed no sianifieant difference with the distribution of the two types of simple infection in the two groups （P〉0.05）. But the rate of mixing infection is much higher in the patients with vaginitis （P〈O. 05）. Conclusion The pathology of Mycoplasma urealyticum may be related to the mixed infection of different biotypes.