Comparison of FISH PRINS, and conventional and fluorescent PCR for single-cell sexing: suitability for preimplantation genetic diagnosis.
OBJECTIVE
Although conventional polymerase chain reaction (PCR) was the first method used for sexing in preimplantation genetic diagnosis, fluorescent in situ hybridization (FISH) has become the method of choice. Recently two new techniques, primed in situ synthesis (PRINS) and fluorescent PCR, have been developed. This study compares the reliability and accuracy of these four techniques in single cells.


RESULTS
In buccal cells, fluorescent PCR and FISH had similar reliability (94 and 93%) and accuracy (97 and 96%) rates. The reliability and accuracy of PRINS (91 and 25%) and conventional PCR (79 and 89%) were lower. In human blastomeres, FISH and fluorescent PCR had similar reliability (100%, 717; 95%, 190/201) rates. Accuracy rates were 71% (517) and 99% (188/190) for FISH and fluorescent PCR, respectively, however, too few blastomeres were analyzed by FISH for meaningful comparison. However, when these data are compared with published data, the method of choice for blastomere sexing appears to be fluorescent PCR.


CONCLUSIONS
Flouroscent PCR has major implications for PGD.
Findlay
I
I
Centre for Reproduction, Growth and Development, Leeds University, UK.
Corby
N
N
Rutherford
A
A
Quirke
P
P
D001710:Biotin / Q000031:analogs & derivatives; D001757:Blastomeres / Q000166:cytology; D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D004076:Digoxigenin / Q000031:analogs & derivatives / Q000378:metabolism; D005260:Female; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008297:Male; D009061:Mouth Mucosa / Q000166:cytology; D009865:Oocytes / Q000166:cytology; D016133:Polymerase Chain Reaction / Q000379:methods*; D019849:Sex Determination Processes
D003856:Deoxyuracil Nucleotides; C070202:digoxigenin-11-deoxyuridine triphosphate; D001710:Biotin; C045931:biotin-11-dUTP; D004076:Digoxigenin