title: Double and triple in situ chromosomal labeling of human spermatozoa by PRINS.
abstract: The PRimed IN Situ (PRINS) labeling method allows rapid, specific detection of human chromosomes in situ. We have adapted the PRINS protocol to mature human sperm in combination with a 3 M NaOH protocol for simultaneous in situ decondensation and denaturation of sperm nuclei. Using fluorochrome-labeled dNTPs in a sequential PRINS reaction, the direct detection of two or three distinct chromosomes must be performed within a timespan of 3 h. The method was tested with primers specific for chromosomes 8, 9, 12, 13, 16, 18, and 21 and the X. The frequencies of disomy ranged from 0.11% to 0.34%. Chromosome-specific primers have been defined for most of the human chromosomes, including some that are indistinguishable by fluorescence in situ hybridization (FISH) with centromeric probes. Consequently, this new strategy constitutes a rapid and efficient alternative to FISH for detecting nondisjunction in human sperm.
authors: CNRS UPR 9008, Montpellier, France.
authors: L
authors: L
authors: Coignet
authors: A
authors: A
authors: Girardet
authors: B
authors: B
authors: Andréo
authors: J P
authors: JP
authors: Charlieu
authors: F
authors: F
authors: Pellestor
mesh_terms: D000782:Aneuploidy; D001483:Base Sequence; D002877:Chromosomes, Human / Q000648:ultrastructure*; D017931:DNA Primers; D004276:DNA, Satellite / Q000032:analysis; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008969:Molecular Sequence Data; D013094:Spermatozoa / Q000648:ultrastructure*
chemical_list: D017931:DNA Primers; D004276:DNA, Satellite