Real-time PCR技术筛查男性不育患者的Y染色体微缺失

Y chromosome micro-deletions in male infertility screened by real-time PCR

摘要:

目的：探讨real-time PCR技术检测Y染色体微缺失的适用性，并研究男性不育患者中无精症和少精症Y染色体微缺失的发生率。方法：本研究收集6例Y染色体微缺失阳性样本，4例女性样本和4例正常生育男性样本分别采用real-time PCR及多重PCR电泳法对Y染色体无精子因子（azoospermia factor，AZF）AZFa、AZFb、AZFc区进行检测；再收集452例临床男性不育患者的外周血样本，采用real-time PCR检测Y染色体微缺失位点。结果：2种方法对14例比对样本6个共有序列标签位点检测的结果一致；452例男性不育症包括96例少弱精症、9例重度少弱精症、69例无精症、3例畸精症和275例其他男性不育症，real-time PCR法共检出8例AZFc区缺失、1例AZFb区缺失和2例AZFbc共缺失，其中在少弱精症、重度少弱精症和无精症中Y染色体微缺失阳性率分别为5.21%、11.1%和7.25%，而另外2组均未检测到微缺失。结论：real-time PCR法检测Y染色体微缺失结果与经典的多重PCR电泳法结果相同，且该法的检测结果与临床诊断相符，因此real-time PCR适用于Y染色体微缺失的临床实验室筛查。

Abstract:

Objectives：To investigate the applicability of real-time PCR in detecting Y chromosome micro-deletions and the incidence of Y chromosome micro-deletions in male infertility due to azoospermia and oligozoospermia. Methods：The specimens，respectively derived from 6 patients with Y chromosome micro-deletions，4 female cases and 4 normal fertile men，were gathered and detected by real-time PCR and multiplex PCR-electrophoresis for Y chromosome azoospermia factor（AZF） AZFa，AZFb，AZFc. Then peripheral blood of 452 male infertile patients were collected，and sequence tagged sites（STSs） of micro-deletions in Y chromosome AZF were detected using real-time PCR. Results：The detection results from the two methods described above respectively，for six consensus STSs of 14 samples，were wholly consistent. In 452 patients with male infertility，including 96 patients with oligo-asthenospermia，9 patients with severe oligozoospermia，69 patients with azoospermia，3 patients with teratospermia and 275 patients with other male in－fertility，eight AZFc region deletions，one AZFb region deletion and two AZFbc region deletions were all found by real time PCR. The positive rate of Y chromosome micro-deletions was respectively 5.21%，11.1% and 7.25% in oligo-asthenospermia，severe oligo－zoospermia and azoospermia，while in other two groups no micro-deletion was detected. Conclusion：Real-time PCR，used to de－tect Y chromosome micro-deletions，has the same results with that of the classical multiplex PCR-electrophoresis，and the test results are consistent with the corresponding clinical diagnosis，therefore real-time PCR is suitable for clinical laboratory screening for Y chromosome micro-deletions.