血清E型沙眼衣原体的Real-time PCR定量分析

Quantitative Analysis of Chlamydia trachomatis (serovar E) by Real-time PCR

目的沙眼衣原体感染是最常见的性传播疾病,本文拟建立一种准确快速、标准化的感染动物组织衣原体载量检测体系。方法体外扩增感染用沙眼衣原体血清E型,克隆衣原体特异基因OMP1基因片段作为标准品,用Real time PCR法测定衣原体基因组拷贝数进行衣原体定量。结果 Real time PCR在OMP1基因片段200至2×108拷贝检测结果成线性,在模板中加入小鼠基因组未出现非特异扩增,同时未影响扩增效率。结论针对衣原体特异基因OMP1的实时定量PCR方法可以较为灵敏的特异的定量检测感染动物样本中的衣原体。

Objective Chlamydia trachomatis infection is the most common sexually transmitted disease.The aim of this research is to set a standard system for determination of chlamydia load in tissues from infected animals.Methods Chlamydia trachomatis（serovar E） was amplified in vitro for infection.The gene fragment of the chlamydia-specific gene OMP1 was cloned as standard.The quantity of chlamydia was determined based on the copy number of chlamydia genome by real time PCR.Results The results of real time PCR were linear when the copy number of OMP1 gene was within the range from 200 to 2 × 10^8.Unspecific amplification was not detected after adding mouse genome into the templates,nor was the amplification efficiency affected.Conclusion Real time PCR targeting chlamydia-specific gene OMP1 could be applied as a standard method to quantitatively analysis chlamydia in infected animal sample with adequate sensitivity and specificity.