MSRE-qPCR法检测少弱精子症患者父源印记基因H19上游区域甲基化水平.docVIP

MSRE-qPCR法检测少弱精子症患者父源印记基因H19上游区域甲基化水平 　　 　　 　　 　　 　　 　　[摘要] 目的 建立精子中父源印记基因H19上游区域MSRE-qPCR检测方法，并分析男性少弱精子症患者精子父源印记基因H19上游区域甲基化水平。 方法 收集20例正常精液样本，同时筛选30例少弱精子症患者精液样本，应用甲基化敏感性限制性内切酶法并结合定量PCR对所有样本父源印记基因H19上游区域甲基化水平进行分析。 结果 正常精液样本父源印记基因H19上游区域平均甲基化率为（99.8±2.72）%，高于少弱精子症患者精液样本的（82.4±15.30）%，差异有统计学意义（P＜0.01）。 结论 MSRE-qPCR法可用于父源印记基因H19上游区域甲基化水平的检测，少弱精子症者精液样本父源印记基因H19上游区域甲基化水平显著低于正常人。 　　[关键词] 印记基因H19；MSRE-qPCR；少弱精子症；DNA甲基化 　　[中图分类号] R698[文献标识码] A[文章编号] 1674-4721（2014）03（c）-0012-04 　　 　　The methylation level detection of the upstream of H19 paternal imprinted gene in olig-asthenospermia patient by MSRE-qPCR 　　YAN Peng1 REN Li-juan2 CHENG Zhen1 GUO Da-wei1▲ BAI Li-juan1 JIA Lin-na1 　　1.Forensic Medicine of Shanxi Medical University,Taiyuan030001,China;2.Shanxi Dayi Hospital,Taiyuan030032,China 　　[Abstract] Objective To establish the methylation level of upstream of H19 paternal imprinted gene in semen of olig-asthenospermia patient and analyse the methylation level detection of the upstream of H19 paternal imprinted gene in olig-asthenospermia patient.Methods The methylation level in the upstream of the H19 paternal imprinted gene of abnormal(30 olig-asthenospermia samples)and normal(20 normal semen samples)was detected respectively by conducting the methylation sensitive restriction endonucleases-quantitative polymerase chain reaction(MSRE-qPCR,asemiquantitative DNA methylation analytical method).Results The average methylation rate in the upstream of the H19 paternal imprinted gene in normal semen samples was(99.8±2.72)%,higher than that in olig-asthenospermia samples(82.4±15.30)%,with statistical difference(P＜0.01).Conclusion The method of MSRE-qPCR can be used in the methylation level detection of upstream of the H19 paternal imprinted gene of semen,and the methylation level of upstream of the H19 paternal imprinted gene in olig-asthenospermia patient is much lower than that in normal person. 　　[Key words] H19 paternal imprinted gene;MSRE-qPCR;Olig-astheno