Martin R H, Rademaker A
Department of Medical Genetics, Faculty of Medicine, University of Calgary, Alberta, Canada.
Mol Reprod Dev. 1995 Sep;42(1):89-93. doi: 10.1002/mrd.1080420112.
Aneuploidy estimates for chromosomes 1, 12, X, and Y were obtained in human sperm from five donors using multicolor fluorescence in situ hybridization (FISH) analysis. Disomy frequencies were obtained by scoring a minimum of 10,000 sperm for each chromosome probe per donor. This analysis was replicated for two scoring criteria: one used one half of a signal domain as the minimum distance between two signals to be counted as two and thus disomic; the other set one signal domain as the minimum distance between two signals. A total of 120,870 sperm were assessed using one half of a domain as the criterion, and 113,478 were scored using one domain as the criterion. The percentage of disomy for chromosomes 1, 12, X, Y, and XY was 0.18, 0.16, 0.15, 0.19, and 0.25, respectively, using the one-half-domain criterion, and 0.08, 0.17, 0.07, 0.12, and 0.16, respectively, using the one-domain criterion. The percentage of disomy decreased significantly with use of one domain as the minimum distance for signal separation for all chromosomes except for chromosome number 12. These lower disomy frequencies correlated well with frequencies derived from human sperm karyotypes analyzed in our laboratory. This suggests that the fluorescent signals for chromosomes 1, X, and Y split into more than one domain in decondensed interphase sperm, and that the use of the one-half-domain criterion would lead to an overestimate of aneuploidy frequencies. The factors known to affect aneuploidy estimates derived from FISH studies are discussed, and recommendations for stringent scoring criteria are proposed.
利用多色荧光原位杂交（FISH）分析，对来自5名供体的人类精子中的1号、12号、X和Y染色体的非整倍体情况进行了评估。通过对每个供体的每个染色体探针至少10,000个精子进行评分来获得二体频率。针对两种评分标准重复了该分析：一种将信号域的一半作为两个信号之间的最小距离，若此距离下两个信号可计为二体；另一种将一个信号域作为两个信号之间的最小距离。以信号域的一半作为标准共评估了120,870个精子，以一个信号域作为标准对113,478个精子进行了评分。使用信号域一半的标准时，1号、12号、X、Y和XY染色体的二体百分比分别为0.18%、0.16%、0.15%、0.19%和0.25%；使用一个信号域的标准时，相应百分比分别为0.08%、0.17%、0.07%、0.12%和0.16%。除12号染色体外，对于所有染色体，使用一个信号域作为信号分离的最小距离时，二体百分比显著降低。这些较低的二体频率与我们实验室分析的人类精子核型得出的频率高度相关。这表明在去浓缩的间期精子中，1号、X和Y染色体的荧光信号会分裂成多个信号域，并且使用信号域一半的标准会导致非整倍体频率的高估。讨论了已知影响FISH研究中非整倍体评估的因素，并提出了严格评分标准的建议。
