生殖支原体TaqMan荧光PCR检测方法的建立

Detection of Mycoplasma genitalium with real-time PCR assay

目的建立一种快速、灵敏、特异的生殖支原体TaqMan荧光PCR检测方法。方法选取生殖支原体mg219基因保守区域设计、合成特异性扩增引物和TaqMan探针,建立并优化荧光PCR检测方法。对优化后的方法进行扩增效率、灵敏度及特异度评价,并与已报道的生殖支原体常规PCR方法进行比较。结果建立的荧光PCR方法对生殖支原体的检测限约为10copies,高于常规PCR的102 copies,并缩短检测时间100min。用该方法扩增20种病原菌染色体及人类染色体,结果均为阴性,特异度为100%。结论建立的荧光PCR方法可快速、灵敏、特异地检测生殖支原体,有望用于临床标本检测。

Objective Established a simple,rapid real-time PCR assay to detect Mycoplasma genitalium.Methods An optimized real-time PCR assay was established by analyzing the mg219 gene of M.genitalium.The specificity and sensitivity of this assay were evaluated and compared with conventional PCR assay using a standard concentration plasmid of M.genitalium.Results The sensitivity of the real-time PCR assay was about 10 copies.The specificity of the assay appeared to be 100%.This real-time PCR assay had about 10 times the sensitivity of conventional PCR.Conclusion This real-time PCR assay maybe a suitable method for the detection of M.genitalium in clinical settings.