Double and triple in situ chromosomal labeling of human spermatozoa by PRINS
- PMID: 8751381
- DOI: 10.1159/000134361
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.
Similar articles
-
Assessment of aneuploidy for chromosomes 8, 9, 13, 16, and 21 in human sperm by using primed in situ labeling technique.Am J Hum Genet. 1996 Apr;58(4):797-802. Am J Hum Genet. 1996. PMID: 8644744 Free PMC article.
-
Rapid chromosome detection by PRINS in human sperm.Am J Med Genet. 2002 Jan 15;107(2):109-14. doi: 10.1002/ajmg.10101. Am J Med Genet. 2002. PMID: 11807883
-
Aneuploidy detection in human sperm nuclei using PRINS technique.Am J Med Genet. 1996 Aug 23;64(3):488-92. doi: 10.1002/(SICI)1096-8628(19960823)64:3<488::AID-AJMG8>3.0.CO;2-L. Am J Med Genet. 1996. PMID: 8862627
-
Detection of chromosomes and estimation of aneuploidy in human spermatozoa using fluorescence in-situ hybridization.Mol Hum Reprod. 1997 Jul;3(7):585-98. doi: 10.1093/molehr/3.7.585. Mol Hum Reprod. 1997. PMID: 9268136 Review.
-
In situ aneuploidy assessment in human sperm: the use of primed in situ and peptide nucleic acid-fluorescence in situ hybridization techniques.Asian J Androl. 2006 Jul;8(4):387-92. doi: 10.1111/j.1745-7262.2006.00137.x. Asian J Androl. 2006. PMID: 16763713 Review.
Cited by
-
Using PRINS for gene mapping in polytene chromosomes.Chromosome Res. 1997 Nov;5(7):463-5. doi: 10.1023/a:1018417013699. Chromosome Res. 1997. PMID: 9421263
-
Fluorescence in situ hybridization with chromosome paint probes: a novel approach to assess aneuploidy in human sperm nuclei.J Assist Reprod Genet. 1999 Jan;16(1):46-51. doi: 10.1023/a:1022549713692. J Assist Reprod Genet. 1999. PMID: 9987694 Free PMC article.