[Comparative studies of serological typing and HLA-A, B antigen genotyping with PCR using sequence-specific primers].
OBJECTIVE
To evaluate the accuracy of PCR with sequence-specific primers (PCR-SSP) for HLA-I genotyping and analyze the causes of the errors occurring in the genotyping.


METHODS
DNA samples and were obtained from 34 clinical patients, and serological typing with monoclonal antibody (mAb) and HLA-A and, B antigen genotyping with PCR-SSP were performed.


RESULTS
HLA-A and, B alleles were successfully typed in 34 clinical samples by mAb and PCR-SSP. No false positive or false negative results were found, and the erroneous and missed diagnosis rates were obviously higher in serological detection, being 23.5% for HLA-A and 26.5% for HLA-B. Error or confusion was more likely to occur in the antigens of A2 and A68, A32 and A33, B5, B60 and B61.


CONCLUSIONS
DNA typing for HLA-I class (A, B antigens) by PCR-SSP has high resolution, high specificity, and good reproducibility, which is more suitable for clinical application than serological typing. PCR-SSP may accurately detect the alleles that are easily missed or mistaken in serological typing.
Wu
Da-lin
DL
Central Laboratory, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. wksys@fimmu.com
Ling
Han-xin
HX
Tang
Hao
H
D000911:Antibodies, Monoclonal / Q000276:immunology; D005838:Genotype; D015234:HLA-A Antigens / Q000145:classification / Q000235:genetics* / Q000276:immunology; D015235:HLA-B Antigens / Q000145:classification / Q000235:genetics* / Q000276:immunology; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D018807:Polymorphism, Single-Stranded Conformational; D012698:Serologic Tests
D000911:Antibodies, Monoclonal; D015234:HLA-A Antigens; D015235:HLA-B Antigens