Cunningham Scott A, Mandrekar Jayawant N, Rosenblatt Jon E, Patel Robin
Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN 55905, USA.
Division of Biomedical Statistics and Informatics, Department of Health Science Research, Mayo Clinic, Rochester, MN 55905, USA.
Int J Bacteriol. 2013;2013:168742. doi: 10.1155/2013/168742. Epub 2013 Mar 11.
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.
目的。我们比较了实验室自行开发的用于检测人型支原体以及用于检测和解脲脲原体与微小脲原体的鉴别并将其与使用提交进行人型支原体和脲原体培养的泌尿生殖系统标本进行培养的方法。方法。对临床细菌学实验室接收的283份用于人型支原体和脲原体培养的泌尿生殖系统标本进行了评估。使用MagNA Pure 2.0上的总核酸试剂盒提取核酸。将5μL提取物与两种主混合液各15μL混合。在LightCycler 480 II系统上进行检测。采用常规方法进行培养。结果。人型支原体PCR检测到42份人型支原体培养阳性标本中的38份,以及2份培养阴性标本(敏感性为90.5%;特异性为99.2%)。脲原体PCR检测到144份脲原体培养阳性标本中的139份,以及9份培养阴性标本(敏感性为96.5%;特异性为93.6%)。在脲原体检测呈阳性的标本中,仅检测到解脲脲原体的有33份,仅检测到微小脲原体的有109份,两者均检测到的有6份。结论。所描述的PCR检测方法是用于检测人型支原体和脲原体的快速替代培养方法,并且与培养不同,脲原体检测方法能够轻松地将解脲脲原体与微小脲原体区分开来。