Wyrobek A J, Robbins W A, Mehraein Y, Pinkel D, Weier H U
Lawrence Livermore National Laboratory, CA 94550.
Am J Med Genet. 1994 Oct 15;53(1):1-7. doi: 10.1002/ajmg.1320530102.
Sex chromosome aneuploidy is the most common numerical chromosomal abnormality in humans at birth and a substantial portion of these abnormalities involve paternal chromosomes. An efficient method is presented for using air-dried smears of human semen to detect the number of X and Y chromosomes in sperm chromatin using two-chromosome fluorescence in situ hybridization. Air-dried semen smears were pre-treated with dithiothreitol and 3,4-diiodosalicylate salt to decondense the sperm chromatin and then were hybridized with repetitive sequence DNA probes that had been generated by PCR and differentially labeled. Hybridizations with X and Y specific probes showed the expected ratio of 50%X:50%Y bearing sperm. Sperm carrying extra fluorescence domains representing disomy for the X or Y chromosomes occurred at frequencies of approximately 4 per 10,000 sperm each. Cells carrying both X and Y fluorescence domains occurred at a frequency of approximately 6/10,000. Thus, the overall frequency of sperm that carried an extra sex chromosome was 1.4/1,000. The frequencies of sperm carrying sex chromosome aneuploidies determined by hybridization did not differ statistically from those reported from the same laboratory using the human-sperm/hamster-egg cytogenetic technique. Multi-chromosome fluorescence in situ hybridization to sperm is a promising method for assessing sex-ratio alterations in human semen and for determining the fraction of sperm carrying sex or other chromosome aneuploidies which may be transmissible to offspring.
性染色体非整倍体是人类出生时最常见的染色体数目异常，其中很大一部分异常涉及父源染色体。本文介绍了一种有效的方法，即使用人类精液的空气干燥涂片，通过双色荧光原位杂交检测精子染色质中X和Y染色体的数量。空气干燥的精液涂片先用二硫苏糖醇和3,4 -二碘水杨酸盐预处理，以使精子染色质解聚，然后与通过聚合酶链反应（PCR）产生并进行差异标记的重复序列DNA探针杂交。与X和Y特异性探针的杂交显示出预期的50%X:50%Y精子比例。携带额外荧光域代表X或Y染色体二体性的精子出现频率约为每10000个精子中有4个。同时携带X和Y荧光域的细胞出现频率约为6/10000。因此，携带额外性染色体的精子总体频率为1.4/1000。通过杂交确定的携带性染色体非整倍体精子的频率与同一实验室使用人精子/仓鼠卵细胞遗传学技术报告的频率在统计学上没有差异。对精子进行多染色体荧光原位杂交是一种有前景的方法，可用于评估人类精液中的性别比例变化，并确定可能传递给后代的携带性染色体或其他染色体非整倍体的精子比例。
