Use of PRINS for preconception screening of polar bodies for common aneuploidies.
It has been shown that preconceptional screening for oocyte aneuploidies could help increase the pregnancy rate after in vitro fertilization (IVF), particularly in cases of advanced maternal age. The FISH (fluorescent in situ hybridization) technique is usually used to examine the first polar body (I-PB) for oocyte screening and so avoid fertilizing and transferring embryos from aneuploid oocytes. We have tested the feasibility of using another technique, the primed in situ (PRINS) reaction for this purpose. PRINS is a rapid, inexpensive method of labelling chromosomes. Chromosomes were labelled by in situ annealing with chromosome-specific oligonucleotide primers, followed by primer extension with labelled nucleotides using Taq DNA polymerase. A total of 183 PRINS reactions were performed with primers for chromosomes 13, 16, 18, 21 or X on 63 I-PBs removed from oocytes that failed to become fertilized during IVF. Each I-PB underwent three successive double-labelling reactions and intense signals were obtained in less than 40 min. Our data suggest that PRINS may be a useful alternative or a complement to FISH for detecting the main aneuploidies in all oocytes obtained after follicular puncture.
Petit
C
C
Centre d'AMP - Maternité Régionale A. Pinard, Laboratoire de Biologie sexuelle, Rue du Dr Heydenreich, BP 4213, 54 042 Nancy Cedex, France. Petit.christophe@wanadoo.fr
Martel-Petit
V
V
Fleurentin
A
A
Monnier-Barbarino
P
P
Jonveaux
P
P
Gerard
H
H
D000328:Adult; D000782:Aneuploidy; D022004:Chromosome Structures; D002875:Chromosomes / Q000648:ultrastructure; D017931:DNA Primers; D004624:Embryo Transfer; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D008856:Microscopy, Fluorescence; D009711:Nucleotides; D009865:Oocytes / Q000648:ultrastructure*; D019836:Preimplantation Diagnosis; D019914:Taq Polymerase
D017931:DNA Primers; D009711:Nucleotides; D019914:Taq Polymerase