RESUMO
In this study we compare the results of HLA-DRB1 genotyping by PCR-RFLP and PCR-MPH. HLA-DR specificities were also performed by LCT. Samples were obtained from 20 Thai patients who were on the waiting list for kidney transplant. DNA was extracted by phenol-chloroform extraction. It was found that the results gave complete agreement with two methods of DNA typing, however, there were 3 discrepancies in assigning serologic DR specificities and DNA subtypes (p = 0.0001) which were due to the cross reactive antibodies and the lack of potent antisera to define proper HLA-DR subtypes by LCT. These PCR techniques can be applied to identify other alleles such as HLA-DPB1 and HLA-DQB1 which will improve the standard histocompatibility testing in the future.
Assuntos
Alelos , Frequência do Gene , Genética Populacional , Genótipo , Antígenos HLA-DR/genética , Teste de Histocompatibilidade , Humanos , Transplante de Rim , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de Restrição , TailândiaRESUMO
Fifty-seven sera obtained from pregnant women who attended antenatal clinic were screened for class I HLA antibodies by standard microlymphocytotoxicity test. The age ranged from 20 to 40 years (mean +/- SD = 30.4 +/- 5). The lymphocytes obtained from 98 HLA-typed persons were used as the panel which would include all common HLA-A and HLA-B antigens and as many rare antigens as possible. Eight out of 57 sera (14%) had HLA antibodies which reacted as follows: 3 sera had nonspecific antibodies, 3 sera had multispecific antibodies (B60 + B48 + B7, B60 + B61 + B13 and B51 + B35 + B53), one serum had a monospecific antibody (A2) and one serum was unidentified. The identified antibodies showed strong reactivity (r > 0.8). These sera will be used as HLA typing sera and exchanged with other HLA laboratories. The screening test for class I HLA antibodies in pregnant women attending the antenatal clinic is useful and practical. Several HLA antisera can be obtained at low cost. The cost of self-screening antisera was US$90/ml while the cost of commercial HLA typing sera is approximately US$250/ml, a factor to be considered by a new HLA laboratory with limited budget.