CAJ | 학술논문

为了建立猪衣原体（Chlamydia suis ）实时荧光定量 PCR（real-time PCR）检测方法，根据猪衣原体（C．cuis ）的 MOMP 基因，选取高度特异保守的区域，设计实时荧光定量 PCR 引物和探针，并构建了重组质粒。结果显示，该方法建立的标准曲线方程为：Y＝-3．2147x＋41．218，线性相关系数为0．9991，扩增效率为104．6％，最低检测量为1．7×102 copies ！μL。该检测方法特异性较好，与鹦鹉热亲衣原体、流产亲衣原体、沙眼衣原体、肺炎嗜衣原体、牛羊亲衣原体、鼠衣原体无交叉反应。批内和批间重复试验的变异系数在0．458％～1．174％范围内，具有较好的重复性；与普通 PCR 方法相比较，该方法具有较高的检出率。建立对猪衣原体的监测和流行病学调查都具有非常重要的意义。
위료건립저의원체（Chlamydia suis ）실시형광정량 PCR（real-time PCR）검측방법，근거저의원체（C．cuis ）적 MOMP 기인，선취고도특이보수적구역，설계실시형광정량 PCR 인물화탐침，병구건료중조질립。결과현시，해방법건립적표준곡선방정위：Y＝-3．2147x＋41．218，선성상관계수위0．9991，확증효솔위104．6％，최저검측량위1．7×102 copies ！μL。해검측방법특이성교호，여앵무열친의원체、유산친의원체、사안의원체、폐염기의원체、우양친의원체、서의원체무교차반응。비내화비간중복시험적변이계수재0．458％～1．174％범위내，구유교호적중복성；여보통 PCR 방법상비교，해방법구유교고적검출솔。건립대저의원체적감측화류행병학조사도구유비상중요적의의。
In order to establish quantitative PCR for detecting Chlamydia suis ,based on the MOMP gene of C .cuis,this study selected specific primers and probe from highly conserved region,the PCR reaction was developed by building the recombinant plasmid.The results showed that the detection limit of the assay was 1.7×102 copies/μL,the calibration curve equation:Y=-3.214 7x+41.218,a correlation coefficient of R2 = 0.999 1,expanding efficiency rate 104.6%.The method used to detect Chlamydophila psittaci , Chlamydophila abortus ,Chlamydia trachomatis ,Chlamydophila pneumoniae ,Chlamydophila pecorum , Chlamydia muridarum had no cross reaction.The real-time PCR was reproducible,the test of its intra-and inter-coefficients of variation from 0.495% to 1.752%.Compared with conventional PCR method,the detection rate of the method established in this study is higher.This method is important for detecting C . cuis and epidemiological investigation.