MOLECULAR ANALYSIS OF HUMAN PLATELET ANTIGEN SYSTEM-1 ANTIGEN ON SINGLE CELLS CAN BE APPLIED TO PREIMPLANTATION GENETIC DIAGNOSIS FOR PREVENTION OF ALLOIMMUNE THROMBOCYTOPENIA

OBJECTIVE: Our purpose was to develop a molecular assay to determine t he human platelet antigen system 1 status on single nucleated cells, i ncluding human blastomeres. STUDY DESIGN: Eighty single cultured lymph oblasts of known human platelet antigen system 1 genotype and 24 media blanks were mixed in blinded fashion. Amplification of a 246 bp deoxy ribonucleic acid fragment and subsequent Nci I restriction digestion w ere performed to distinguish human platelet antigen system 1 a from 1 b alleles. Specificity and sensitivity of the technique were determine d. Eight blastomeres were also tested. RESULTS: Deoxyribonucleic acid amplification at the human platelet antigen system 1 locus was success ful in 95% of the reactions. No media blanks showed amplified deoxyrib onucleic acid. The diagnosis was correct in all homozygous human plate let antigen system 1a or 1b cells; three of 23 heterozygous cells ampl ified but failed to digest with Nci I. Overall specificity was 95%. Al l blastomeres successfully amplified. CONCLUSIONS: The human platelet antigen system 1 status determination is reliable from a single cell a nd can be used for preimplantation genetic diagnosis for the preventio n of alloimmune thrombocytopenia.