Department of Pathology and Laboratory Medicine, University of California Los Angeles, Los Angeles, California.
Department of Pathology and Laboratory Medicine, University of California Los Angeles, Los Angeles, California.
J Mol Diagn. 2023 Nov;25(11):838-848. doi: 10.1016/j.jmoldx.2023.07.004. Epub 2023 Sep 7.
Bacterial commensals of the human genitourinary tract, Mycoplasma hominis and Ureaplasma species (parvum and urealyticum) can be sexually transmitted, and may cause nongonococcal urethritis, pelvic inflammatory disease, and infertility. Mycoplasma hominis and Ureaplasma species may also cause severe invasive infections in immunocompromised patients. Current culture-based methods for Mycoplasma/Ureaplasma identification are costly and laborious, with a turnaround time between 1 and 2 weeks. We developed a high-throughput, real-time multiplex PCR assay for the rapid detection of M. hominis and Ureaplasma species in urine, genital swab, body fluid, and tissue. In total, 282 specimens were tested by PCR and compared with historic culture results; a molecular reference method was used to moderate discrepancies. Overall result agreement was 99% for M. hominis (97% positive percentage agreement and 100% negative percentage agreement) and 96% for Ureaplasma species (96% positive percentage agreement and 97% negative percentage agreement). Specimen stability was validated for up to 7 days at room temperature. This multiplex molecular assay was designed for implementation in a high-complexity clinical microbiology laboratory. With this method, >90 samples can be tested in one run, with a turnaround time of 4 to 5 hours from specimen extraction to reporting of results. This PCR test is also more labor effective and cheaper than the conventional culture-based test, thus improving laboratory efficiency and alleviating labor shortages.
人体泌尿生殖道的共生细菌，人型支原体和脲原体（微小脲原体和脲解支原体）可通过性传播，可能导致非淋菌性尿道炎、盆腔炎和不孕。人型支原体和脲原体也可能导致免疫功能低下患者发生严重侵袭性感染。目前基于培养的方法用于鉴定支原体/脲原体既昂贵又繁琐，周转时间为 1 至 2 周。我们开发了一种高通量、实时多重 PCR 检测方法，用于快速检测尿液、生殖器拭子、体液和组织中的人型支原体和脲原体。总共用 PCR 检测了 282 个标本，并与历史培养结果进行了比较；使用分子参考方法来调和差异。对于人型支原体，总体结果一致性为 99%（97%的阳性百分比一致性和 100%的阴性百分比一致性），对于脲原体，总体结果一致性为 96%（96%的阳性百分比一致性和 97%的阴性百分比一致性）。标本稳定性在室温下可验证长达 7 天。该多重分子检测方法设计用于在高复杂度临床微生物学实验室中实施。使用这种方法，一个运行可以测试超过 90 个样本，从标本提取到报告结果的周转时间为 4 至 5 小时。与传统的基于培养的检测方法相比，该 PCR 检测方法更节省劳动力且成本更低，从而提高了实验室效率并缓解了劳动力短缺。
