Detection of aneuploidy in human interphase spermatozoa by fluorescence in situ hybridization (FISH).
Fluorescence in situ hybridization (FISH) with repetitive-sequence DNA probes was performed on human interphase sperm to determine the utility of this technique for aneuploidy detection. DNA sequences specific for chromosomes 15 and 16 and the Y were biotinylated and hybridized with human sperm that had been treated with cetyltrimethylammonium bromide and dithiothreitol to render them accessible to the probes. Fluoresceinated avidin and antiavidin were utilized to visualize bound probe. More than 10,000 sperm were analyzed for each chromosome probe. The hybridization efficiency was 98-99% for all three probes. The frequencies for disomy (i.e., for nuclei containing two fluorescent signals) for chromosomes 15 and 16 and the Y were 0.14%, 0.17%, and 0.11%, respectively. These frequencies are all somewhat higher than those obtained by the human sperm karyotyping technique. Nevertheless, these preliminary results suggest that FISH is a simple and rapid technique that may provide an accurate screen for aneuploidy detection in human sperm.
Department of Pediatrics, Faculty of Medicine, University of Calgary, Alberta, Canada.
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D000782:Aneuploidy; D002884:Chromosomes, Human, Pair 15; D002885:Chromosomes, Human, Pair 16; D005069:Evaluation Studies as Topic; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D013094:Spermatozoa / Q000648:ultrastructure; D014998:Y Chromosome