S. Cunningham, J. Mandrekar, J. Rosenblatt, Robin Patel
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引用次数: 55
摘要
目标。我们将实验室开发的用于人支原体检测、解脲支原体和细小体检测和分化的实时PCR方法与用于人支原体和脲支原体培养的泌尿生殖道标本进行了比较。方法:对临床细菌学实验室收集的283份泌尿生殖系统标本进行人支原体和脲支原体培养。使用MagNA Pure 2.0上的总核酸试剂盒提取核酸。提取液5 μL与两种主混合物各15 μL复配。在LightCycler 480 II系统上进行检测。采用常规方法进行培养。结果。人支原体PCR检测到42例人支原体培养阳性标本中有38例,培养阴性标本中有2例(灵敏度为90.5%;特异性,99.2%)。脲原体PCR检出培养阳性139/144例,培养阴性9例(灵敏度96.5%;特异性,93.6%)。在脲原体检测呈阳性的标本中,33例单独检出解脲原体,109例单独检出细小脲原体,6例两者均检出。结论。所描述的PCR检测是快速替代培养检测人支原体和脲原体的方法,而且,与培养不同,脲原体检测很容易区分解脲原体和细小体。
Rapid PCR Detection of Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum
Objective. We compared laboratory developed real-time PCR assays for detection of Mycoplasma hominis and for detection and differentiation of Ureaplasma urealyticum and parvum to culture using genitourinary specimens submitted for M. hominis and Ureaplasma culture. Methods. 283 genitourinary specimens received in the clinical bacteriology laboratory for M. hominis and Ureaplasma species culture were evaluated. Nucleic acids were extracted using the Total Nucleic Acid Kit on the MagNA Pure 2.0. 5 μL of the extracts were combined with 15 μL of each of the two master mixes. Assays were performed on the LightCycler 480 II system. Culture was performed using routine methods. Results. M. hominis PCR detected 38/42 M. hominis culture-positive specimens, as well as 2 that were culture negative (sensitivity, 90.5%; specificity, 99.2%). Ureaplasma PCR detected 139/144 Ureaplasma culture-positive specimens, as well as 9 that were culture negative (sensitivity, 96.5%; specificity, 93.6%). Of the specimens that tested positive for Ureaplasma species, U. urealyticum alone was detected in 33, U. parvum alone in 109, and both in 6. Conclusion. The described PCR assays are rapid alternatives to culture for detection of M. hominis and Ureaplasma species, and, unlike culture, the Ureaplasma assay easily distinguishes U. urealyticum from parvum.