多重荧光PCR在Y染色体微缺失检测中的应用  

Multiplex fluorescence PCR for detection of Y Chromesome microdeletions

作　　者：李卫巍[1] 韩莹 殷玉玲 吴秋月[1] 张静[1] 蒋卫军[1] 夏欣一[1] LI Wei-wei;HAN Ying;YIN Yu-ling;WU Qiu-yue;ZHANG Jing;JIANG Wei-jun;XIA Xin-yi(PLA Research Institute of Clinical Laboratory Medicine,Nanjing General Hospital of Nanjing Military Region,Nanjing,Jiangsu 210002,China)

机构地区：[1]南京军区南京总医院解放军临床检验医学研究所,江苏南京210002

出　　处：《中华男科学杂志》2018年第11期1011-1015,共5页National Journal of Andrology

基　　金：江苏省科技计划项目(BE2018713);南京市科技计划项目(201605004);国家人口计生委重点实验室开放基金课题(yjjc201501)~~

摘　　要：目的:Y染色体AZF微缺失检测是国内外公认的男性不育辅助检查之一,传统的PCR凝胶电泳因其缺点已不太适应临床需要。本文旨在探讨多重荧光PCR在AZF微缺失检测中的可行性。方法:收集AZF微缺失各种类型样本238例,正常男性样本62例,比对两种不同的检测方法(多重PCR凝胶电泳和多重荧光PCR),Kappa检验比较分析两种方法检测结果的一致性。结果:多重PCR凝胶电泳和多重荧光PCR检测300例临床标本,两者在位点检出率方面完全一致,差异无统计学意义。Kappa检验显示Y染色体AZFa、AZFb、AZFc 3个区6个位点P值及Kappa值均为1,两种方法一致性好,无统计学差异。结论:多重荧光PCR技术比多重PCR凝胶电泳法可大量节约时间,减少工作量,提高临床检验工作效率,是临床Y染色体微缺失检测的优选方法。Objective： Detection of azoospermia factor（ AZF） microdeletions on the Y chromosome is one of the auxiliary strategies recognized at home and abroad for the examination of male infertility. Traditional PCR gel electrophoresis fails to meet the clinical needs due to its shortcomings. The purpose of this study was to explore the feasibility of multiplex fluorescence PCR in the detection of AZF microdeletions. Methods： We collected samples of Y chromosomal AZF microdeletions from 238 patients with azoospermia or oligozoospermia and 62 normal males,identified the 14 short tandem repeat（ STR） loci in the AZF region of the Y chromosome by multiplex PCR gel electrophoresis and multiplex fluorescence PCR,and analyzed the consistency in the results of the two methods by Kappa test. Results： There was a perfect consistency between multiplex PCR gel electrophoresis and multiplex fluorescence PCR in the detection rate of the STR loci in the 300 samples. Kappa test showed both P and Kappa values to be 1 for the 6 loci in the AZFa,AZFb and AZFc regions of the Y chromosome,with no statistically significant difference between the two methods. Conclusion： Multiplex fluorescence PCR can save a lot of time,reduce workload and improve laboratory efficiency and therefore is preferable to multiplex PCR gel electrophoresis in detecting Y chromosome microdeletions.

关 键 词：Y染色体 AZF微缺失 多重PCR凝胶电泳 多重荧光PCR 

分 类 号：R446.9[医药卫生—诊断学]