mRNA荧光复合扩增系统鉴别正常和异常精液初探  

Identify normal and abnormal semen via m RNA fluorescence multiplex amplification system: A preliminary study

作　　者：王冲[1] 赵禾苗[1,2] 林清峦 胡兰[1] 畅晶晶[1] 凃政[1] 

机构地区：[1]公安部物证鉴定中心法医遗传学公安部重点实验室,北京100038 [2]西安交通大学,陕西西安710049 [3]中国人民公安大学,北京100038 [4]海南省琼海市公安局,海南琼海571400

出　　处：《中国法医学杂志》2017年第5期453-456,461,共5页Chinese Journal of Forensic Medicine

基　　金：公安部技术研究计划重点项目(2015JSYJA02)

摘　　要：目的构建mRNA荧光复合扩增体系,实现对不同种类精液(尤其是无精症精液)的区分鉴别。方法收集正常、少精症及无精症的精液样本,制备精斑样本后提取细胞总RNA,利用逆转录PCR技术扩增2个精子特异mRNA标记(PRM1、PRM2)、2个精浆特异mRNA标记(TGM4、SEMG1)和2个管家基因mRNA标记(TEF、UCE)。结果正常精液样本可检测到全部精液mRNA标记表达;少精症精液样本虽然可检测到全部mRNA标记表达,但精子特异mRNA标记表达量较低;无精症精液样本不能检测到精子mRNA标记,只能检测到精浆特异mRNA标记。结论利用mRNA荧光复合扩增系统可以实现对正常和无精症精液的区分,而正常精液和少精症精液相比差异无统计学意义。Objective Construct a m RNA multiplex amplification system to identify different types of semen stains. Methods First, collect normal, oligozoospermia and azoospermia semen samples to make semen stains. Second, extract total RNA with Qiagen RNeasy Micro Kit. Then use reverse transcript PCR to amplify goal m RNA markers： 2 markers for sperm（PRM1, PRM2）, 2 markers for seminal plasma（TGM4, SEMG1） and 2 housekeeping genes（TEF, UCE）. Results All semen m RNA markers can be detected in normal semen samples. The RFU of sperm m RNA markers are lower in oligozoospermia semen samples than that in normal controls. No sperm m RNA markers can be detected in the azoospermia semen samples, only seminal plasma specific can be detected. Conclusion The differentiation of normal and azoospermia semen can be achieved by using multiplex m RNA fluorescence amplification system. While normal semen and oligozoospermia semen compared to no statistical difference.

关 键 词：法医物证学 MRNA 体液斑迹 精液 无精子症 

分 类 号：D919.4[医药卫生—法医学]