荧光原位杂交(FISH)检测人精子染色体非整倍体率方法的研究  

STUDY ON THE ANEUPLOIDY FREQUENCY IN HUMAN SPERM CHROMOSOME USING FLUORESCENCE IN SITU HYBRIDIZATION (FISH)

作　　者：郑履康[1] 邓丽霞[1] 张桥[1] 

机构地区：[1]中山医科大学公共卫生学院遗传毒理研究室

出　　处：《癌变．畸变．突变》1998年第5期267-272,共6页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：自然科学基金;中华医学基金

摘　　要：本研究建立了荧光原位杂交（ＦＩＳＨ）方法，并测定了十名正常健康男子精子Ｘ染色体非整倍体率。取少量精液经洗后制片，用二梳苏糖醇（ＤＴＴ）和二碘水杨酸锂（ＬＩＳ）处理，使精子头部紧密的染色质去凝集，然后与生物素标记的α－卫星Ｘ染色体特异ＤＮＡ探针（ＤＸＺ１）作原位杂交；用Ｃｙ３－链亲和素检测杂交信号。结果在荧光显微镜下可清楚见到带有红色杂交点信号的Ｘ精子。杂交率平均为４８６２％（４７５４％－４９３６％）。此值与Ｘ精子占５０％的理论值差别无显著性。总共计数５５４５３个Ｘ精子，Ｘ染色体双体率为００６％；此结果与国外一些学者先前报导的结果一致，本研究提供的正常人Ｘ精子双体率的资料。The frequency of Human sperm chromosome aneuploidy was assessed from 10 normal and health adults by using the method of FISH. Sperm heads were decondensed with dithilthreitol (DTT) and lithium diiodosalicylate (LIS), then the sperm nuclei were hybridized with biotin labeled alpha satellite X chromosome specific DNA probe (DXZ1) in 37℃ overnight. The hybridization signals were detected with Cy3 Streptavidin. The red hybridization signals in X sperms were clearly visible under the Nikon fluorescence microscope. The hybridization efficiency was 48.62% (rance 47.54% to 49.36%). A total 55453 X sperm nuclei were observed.The frequency of disomic sperms was 0.06%. There was no significant difference observed in disomic frequency of X sperms between ours and that reported by former authors. The information provided by this paper is valuable for further studies on men exposed occupationally or environmentally to potential aneuploidogenic agents.

关 键 词：荧光原位杂交 人精子染色体 非整倍体率 

分 类 号：R446.1[医药卫生—诊断学]