摘要
目的建立检测Y染色体无精子症因子(AZF)微缺失的多重定量荧光PCR体系。方法以5′FAM、JOE和TAMRA荧光基团标记PCR引物,建立包含Y染色体AZF 4个亚区(AZFa^d)15个序列标签位点(STS)的多重定量荧光PCR体系;并对无精子症组、严重少精子症组及精液正常组进行Y染色体AZF微缺失检测。结果成功建立了检测Y染色体AZF微缺失的多重定量荧光PCR体系;200例男性中检测到Y染色体AZF微缺失16例,其中72例无精子症组7例,缺失率为9.7%,78例严重少精子症组9例,缺失率为15.4%,50例精液正常组未检测到缺失。无精子症组、严重少精子症组缺失率与精液正常组比较差异均有统计学意义(P<0.05)。结论多重定量荧光PCR技术是一种快速、简便检测Y染色体AZF微缺失的方法,具有重要临床应用价值。
Objective To set up a multiplex quantitative fluorescent PCR system for the detection of azoospermia factor （AZF） microdeletion on Y chromosome. Methods For each pair of primers, the 5＇end of either forward or reverse primer was labeled with a 5 FAM,JOE or TAMRA fluorescent dye to establish multiplex quantitative fluorescent PCR system specific for 15 sequence-tagged sites in 4 subregions （including AZFa-d） of AZF on Y chromosome. The detection of Y chromosome AZF microdeletion was carried out on azoospermia group, oligzoospermia group and normal group. Results A multiplex quantitative fluorescent PCR system was set up for the detection of Y chromosome AZF microdeletion. 16 cases of Y chromosome AZF microdeletion were found in 200 male subjects.7 cases （9.7%） were found in 72 patients in the azoospermia group.And 9 cases （15.4%） were found in 78 patients in the oligzoospermic group. Microdeletion was not found in 50 normal subjects. Differences of deletion rates of azoospermia group,oligozoospermia group and normal group were statistically significant differences（P〈0.05）. Conclusion Multiplex quantitative fluorescent PCR is a simple method for the clinical detection of Y chromosome microdeletion.
出处
《热带医学杂志》
CAS
2014年第1期26-31,87,共7页
Journal of Tropical Medicine
基金
广东省科技计划项目(2010B031600120)
关键词
多重定量荧光PCR
Y染色体
无精子症因子
multiplex quantitative fluorescent PCR
Y chromosome
azoospermia factor
