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 the human platelet antigen system 1 status on single nucleated cells, including human blastomeres.


METHODS
Eighty single cultured lymphoblasts of known human platelet antigen system 1 genotype and 24 media blanks were mixed in blinded fashion. Amplification of a 246 bp deoxyribonucleic acid fragment and subsequent Nci I restriction digestion were performed to distinguish human platelet antigen system 1a from 1b alleles. Specificity and sensitivity of the technique were determined. Eight blastomeres were also tested.


RESULTS
Deoxyribonucleic acid amplification at the human platelet antigen system 1 locus was successful in 95% of the reactions. No media blanks showed amplified deoxyribonucleic acid. The diagnosis was correct in all homozygous human platelet antigen system 1a or 1b cells; three of 23 heterozygous cells amplified but failed to digest with Nci I. Overall specificity was 95%. All blastomeres successfully amplified.


CONCLUSIONS
The human platelet antigen system 1 status determination is reliable from a single cell and can be used for preimplantation genetic diagnosis for the prevention of alloimmune thrombocytopenia.
Van den Veyver
I B
IB
Institute for Molecular Genetics, Baylor College of Medicine, Houston, TX 77030.
Chong
S S
SS
Kristjansson
K
K
Snabes
M C
MC
Moise
K J
KJ
Hughes
M R
MR
D016824:Antigens, Human Platelet / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D002886:Chromosomes, Human, Pair 17; D017931:DNA Primers; D005260:Female; D005315:Fetal Diseases / Q000175:diagnosis* / Q000276:immunology / Q000517:prevention & control; D005817:Genetic Counseling; D005838:Genotype; D006801:Humans; D039661:Integrin beta3; D007518:Isoantibodies / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D011696:Purpura, Thrombocytopenic / Q000175:diagnosis* / Q000235:genetics / Q000276:immunology / Q000517:prevention & control; D012680:Sensitivity and Specificity
D016824:Antigens, Human Platelet; D017931:DNA Primers; C061248:ITGB3 protein, human; D039661:Integrin beta3; D007518:Isoantibodies; C083451:human platelet antigen 1b