DETECTION OF ANEUPLOIDY IN HUMAN SPERMATOZOA USING FLUORESCENCE IN-SITU HYBRIDIZATION (FISH)

Fluorescence in situ hybridization (FISH) with chromosome specific alp ha-satellite DNA probes was used to estimate the rates of aneuploidy o f chromosomes 1, 17, 18, X and Y in human ejaculated sperm. Sperm samp les were collected from six donors, and biotinylated DNA probes, D1Z5, D17Z1, D18Z1, DXZ1 and DYZ3 were hybridized to interphase sperm which had been pretreated with dithiothreitol to expand their nuclei. A min imum of 3,000 sperm per donor were analyzed. The hybridization efficie ncy was 99.68% for all the five probes. The frequencies of aneuploidy for chromosomes 1, 17 and 18 were 0.65%, 0.66%, and 0.61%, respectivel y. For XX- and YY-sperm the frequencies were 0.28% and 0.27%, respecti vely. To estimate the diploidy and disomy rates, a mixture of D17Z1 an d D18Z1 were used as probes, and the frequency of diploid sperm was ca lculated to be 0.27%. After subtraction of the diploidy rate, the diso my rates for chromosomes 1, 17, and 18 were estimated to be 0.38%, 0.3 9% and 0.33%, respectively. The proportion of X- and Y-bearing sperm w ere 49.90% and 49.66%, consistent with an expected 1:1 ratio.