Evaluation of PCR for the Identification of Mycoplasma spp. and Mycoplasma genitalium congenital infection in cord blood

Sakineh Monzavi, M. Shahhosseiny, Mahsa Malekmohammadi Kalahroudi
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Abstract

Background: Congenital infections are the most important threat to human fetal health caused by bacterial, viral, and fungal agents. In this study, PCR diagnostic method has been used on umbilical cord samples to diagnose bacterial agents such as Mycoplasmas, especially Mycoplasma genitalium. Materials and Methods: This study was performed on 100 cord blood samples from cesarean section patients. Limit of detection (LOD) and specificity tests of both PCR was performed. DNA samples were extracted by the DNG plus method and amplified by the PCR technique. The optimized PCR product of 715 bp for Mycoplasma spp. and 427 bp for M. genitalium was amplified and observed on 1.5% gel electrophoresis. DNA specificity testing of seven other organisms revealed a 100% specificity of these primers. The detection limit was set at 100 copy / reaction for Mycoplasma.spp and 1000 copy / reaction for M. genitalium. Results: Of the 100 samples examined, 5 samples were positive for Mycoplasma spp, and no positive cases were observed for M. genitalium. Maternal infections can have important consequences on the fetus. Conclusion: Molecular methods such as PCR could be used for rapid identification of important factors in congenital infections, such as mycoplasma, and provide a good prognosis.
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脐带血支原体和生殖道支原体先天性感染的PCR鉴定评价
背景:由细菌、病毒和真菌引起的先天性感染是对人类胎儿健康最重要的威胁。本研究采用PCR诊断方法对脐带标本进行支原体,特别是生殖道支原体等细菌病原体的诊断。材料与方法:本研究采用100例剖宫产患者的脐带血样本。进行两种PCR的检出限(LOD)和特异性试验。DNA样品采用DNG +法提取,PCR技术扩增。优化后的支原体和生殖支原体PCR扩增产物分别为715 bp和427 bp,经1.5%凝胶电泳观察。另外7种生物的DNA特异性测试显示,这些引物的特异性为100%。支原体的检出限为100拷贝/反应。生殖支原体1000拷贝/反应。结果:100份标本中支原体阳性5份,生殖支原体阳性未见一例。母体感染会对胎儿造成严重影响。结论:PCR等分子方法可快速鉴定支原体等先天性感染的重要因素,预后良好。
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