Rapid PCR Detection of Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum.
Rapid PCR Detection of Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum.
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DOI：
10.1155/2013/168742
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
Patel R
中科院分区：
文献类型：
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作者：
Cunningham SA;Mandrekar JN;Rosenblatt JE;Patel R
目标。我们比较了实验室开发的检测人型支原体以及检测和区分解脲支原体和细小支原体的实时荧光聚合酶链式反应方法与提交给人支原体和解脲支原体培养的泌尿生殖道标本的培养。方法：研究方法。对临床细菌学实验室采集的283份泌尿生殖道标本进行人型支原体和解脲支原体培养。用Magna Pure 2.0上的总核酸试剂盒提取核酸。将提取物的5 μL与两种母液各15 μL组合。检测在LightCycler 480 II系统上进行。用常规方法进行培养。结果： 人型支原体聚合酶链式反应检测出38/42例人型支原体培养阳性标本，以及2例培养阴性标本(敏感性，90.5%；特异性，99.2%)。在144例解脲支原体培养阳性标本中检出139例，培养阴性9例(敏感性96.5%，特异性93.6%)。在解脲支原体检测阳性的标本中，单独检出解脲支原体33例，单独检出解脲支原体109例，同时检出两种解脲支原体6例。在检测人型支原体和解脲支原体方面，所描述的聚合酶链式反应是快速替代培养的方法，而且，与培养不同的是，解脲支原体检测很容易区分解脲支原体和细小支原体。
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.