【摘要】 目的通过遗传物理图谱对Y染色体大片段缺失的断裂位点进行精确的定位,研究Y染色体无精子症因子(azoospermia factor,AZF)区域微缺失与无精症的关系。方法应用多重PCR在4组反应管中对AZFa区的序列标签位点(sequence tagged site,STS)即sY82,sY84,sY86,AZFb区的sY124,sY127,sY128,sY133,sY134,sY143,AZFc区的sY239,sY242,sY254,sY255和AZFd区的sY145,sY152共15个STS位点进行扩增,以性别决定因子为内控,根据多重PCR结果对sY82,sY86,sY85,sY84进行单独扩增。结果对照组中的15个STS位点及单独扩增的sY85中均有特异性扩增产物,而Y染色体异常患者样品仅有sY82,sY86扩增产物,其余呈阴性。从而将患者的近着丝粒端的断裂位点定位于sY86与sY85之间。结论本研究为该患者的Y染色体大片段缺失断裂位点的精确定位提供了直接的分子生物学证据,建立了近着丝粒端的缺失图谱,证明了该患者无精的原因为AZF基因缺失。更多还原

【Abstract】 Objective To elucidate the relationship between azoospermia factor(AZF) microdeletion of Y chromosome and azoospermia, the exact breakpoint of a severe Y-chromosome deletion was determined according to the physical map of AZF. Methods Multiplex polymerase chain reaction was used to amplify fifteen sequence tagged sites (STS), namely sY82, sY84, sY86 in AZFa, sY124, sY127, sY128, sY133, sY134, sY143 in AZFb, sY239, sY242 sY254, sY255 in AZFc, and sY145, sY152 in AZFd; sex-determing region Y(SRY) was taken as an internal control. And then sY82,sY86,sY85,sY84 were further analyzed using the sample of the patient who had Y-chromosome deletion by G band anlysis to map the breakpoint at molecular level. Results All 15 STS and sY85 were amplified in positive control while only sY82, sY86 were amplified in the clinical sample, thus the breakpoint was found to be between sY86 and sY85. Conclusion This study on the patient provided the direct biomolecular evidence of the exact breakpoint of the severe Y-chromosome deletion and established the deletion map of acrocentric chromosome. It also proved that the patient’s azoospermia was due to the deletion of AZF.更多还原