Analysis of HLA-DRB1*-A* and -B* alleles in prenatal diagnosis for determination of maternal contamination in fetal DNA.
During chorionic villi sampling for prenatal diagnosis with molecular biology techniques, contamination by maternal decidua frequently occurs and can lead to misinterpretation of the test results. To avoid such problems, we present a new method for appraising maternal contamination of fetal DNA, based on genomic typing of the highly variable human leukocyte antigen (HLA) locus-DRB1*, locus A* and locus B* regions by genetic amplification with sequence-specific primers and PCR. Fetal DNA samples obtained for beta-thalassemia diagnosis were analysed after artificial contamination with increasing maternal DNA concentrations ranging from 0.5 to 10% (0.5, 1, 3, 5 and 10%). The approach was found to be rapid, specific, reproducible and highly sensitive and permits recognition of 1-3% contamination by maternal DNA concentrations. The system currently used for detecting maternal DNA contamination in fetal samples is the analysis of polymorphic loci by variable number of tandem repeats and/or short tandem repeats. We propose that the analysis of HLA alleles may provide a valid alternative or complement to this system.
Pietrapertosa
A
A
Division of Hematology II, University of Bari, Piazza Giulio Cesare 11, 70124, Italy. ematolog@cimedoc.uniba.it
Campanale
D
D
Palma
A
A
Renni
R
R
D'Addario
V
V
Tannoia
N
N
Vitucci
A
A
D000483:Alleles; D004247:DNA / Q000032:analysis*; D005260:Female; D005333:Fetus / Q000502:physiology*; D005838:Genotype; D006684:HLA-DR Antigens / Q000032:analysis / Q000235:genetics*; D059811:HLA-DRB1 Chains; D006801:Humans; D011247:Pregnancy; D011296:Prenatal Diagnosis
D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains; D004247:DNA