Dutro Susan M, Hebb Jennifer K, Garin Cresley A, Hughes James P, Kenny George E, Totten Patricia A
Department of Medicine, University of Washington, Seattle, USA.
Sex Transm Dis. 2003 Oct;30(10):756-63. doi: 10.1097/01.OLQ.0000078821.27933.88.
Mycoplasma genitalium is associated with, and could be the cause of, idiopathic cases of urethritis, endometritis, and cervicitis. Further epidemiologic studies on this organism are needed, but currently used polymerase chain reaction (PCR) assays are labor-intensive and culture is insensitive.
The goal was to develop and evaluate a microwell-plate-based PCR assay for M. genitalium.
We adapted an M. genitalium PCR assay targeting the MgPa gene to a 96-microwell plate format with colorimetric detection of PCR products and incorporation of an internal inhibition control to determine the limit of detection of this assay (termed MgPa-IMW) for M. genitalium DNA and evaluate its performance on cervical and male urine specimens.
The MgPa-IMW PCR assay detected 1 and 17 genome copies of M. genitalium (with 27% and 95% confidence) and was able to detect specimens inhibited for amplification. This assay was 100% concordant (50 positive and 50 negative) with the Southern-blot-based PCR assay with cervical specimens. Similarly, this test was 89% concordant with the Southern-blot-based assay for 64 male urine specimens (25 positive, 32 negative, 7 discordant), 97% concordant after correcting for specimens no longer positive by the Southern blot-based assay after freezer storage.
The MgPa-IMW assay is sensitive and specific for the detection of M. genitalium in patient specimens and should facilitate large-scale screening for this organism.
生殖支原体与特发性尿道炎、子宫内膜炎和宫颈炎有关，可能是其病因。需要对这种微生物进行进一步的流行病学研究，但目前使用的聚合酶链反应（PCR）检测方法劳动强度大，且培养法不敏感。
开发并评估一种基于微孔板的生殖支原体PCR检测方法。
我们将一种针对MgPa基因的生殖支原体PCR检测方法改编为96孔板形式，采用比色法检测PCR产物，并加入内部抑制对照，以确定该检测方法（称为MgPa-IMW）对生殖支原体DNA的检测限，并评估其在宫颈和男性尿液标本中的性能。
MgPa-IMW PCR检测方法能检测到1个和17个生殖支原体基因组拷贝（置信度分别为27%和95%），并能检测到被抑制扩增的标本。该检测方法与基于Southern印迹的宫颈标本PCR检测方法100%一致（50个阳性和50个阴性）。同样，对于64份男性尿液标本（25个阳性、32个阴性、7个不一致），该检测方法与基于Southern印迹的检测方法一致性为89%，在对冷冻保存后经Southern印迹检测不再呈阳性的标本进行校正后，一致性为97%。
MgPa-IMW检测方法对患者标本中生殖支原体的检测具有敏感性和特异性，应有助于对这种微生物进行大规模筛查。
