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

The methylation level detection of the upstream of H19 paternal imprinted gene in olig-asthenospermia patient by MSRE-qPCR

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。

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 ab- normal（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,asemiquantita- tire DNA methylation analytical method）.Results The average methylation rate in the upstream of the H19 paternal im- printed gene in normal semen samples was（99.8±2.72）%,higher than that in olig-asthenospermia samples（82.4±15.30）%, with statistical differenee（P〈0.01）.Conclusion The method of MSRE-qPCR can be used in the methylation level detec- tion of upstream of the H19 paternal imprinted gene of semen,and the methylation level of upstream of the H19 pater- nal imprinted gene in olig-asthenospermia patient is much lower than that in normal person.