SYBR荧光定量方法检测宫颈脲原体在妇产科中的应用

Application of cervix ureaplasma detected with SYBR fluorescent quantitation method in gynaecology and obstetrics department

目的建立基于SYBR的快速检测Ureaplasma urealyticum（UU）和Ureaplasmaparvum（UP）的荧光定量检测方法。方法根据UU、UP的基因序列,设计分别针对UU、UP的特异性引物,建立了它们的荧光定量检测体系。通过Tm值和对其他病原菌的分析验证其特异性,应用于临床样品验证其实用性。结果在1×10^1～1×10^8拷贝/2μL DNA范围内拷贝数与CT值呈现良好的线性关系,检测灵敏度1×10^1～1×10^9拷贝/2μL DNA,重复3次的相关系数为0.65%～1.75%,重复性好。结论建立的荧光定量检测体系能够快速、简便地进行UU、UP的分型鉴定,具有实际应用价值。

Objective It is to establish the fluorescent quantitation method which is based on SYBR and quickly detecting Ureaplasma urealyticum （UU） and Ureaplasma parvum （UP）. Methods According the gene order of UU and UP, specificity primer which was aimed directly at UU and UP respectively was designed. Their fluorescent quantitation detecting system was established. The specificity was validated through Tm value and analyzing others pathogenic bacteria. The practicality was validated through applying clinical specimen. Results The copy number had better linear correlation with CT value in the extent from 1×10^1 to 1×10^8 copies/2μL DNA. The detectability was from 1×10^1 to 1×10^9 copies/2μL DNA. The correlation coefficient which was repeated for three times was 0.65 % - 1.75 %. The repeatability was good. Conclusion The fluorescent quantitation detecting system which is established can quickly and conveniently typing authentication UU and UP, which has practical application value.