HLA-DR typing by polymerase chain reaction amplification with sequence-specific primers in paternity disputes.
BACKGROUND
HLA-DR typing was generally performed by serology before. HLA-DRB1 typing can be achieved by polymerase chain reaction amplification with sequence specific primers (PCR-SSP).


METHODS
In this study, primers for "low-resolution" HLA-DR typing by PCR-SSP were synthesized upon our request by a company in Taiwan. Twenty-eight DNA samples from international standardized DNA Reference Panel and 20 DNA samples with known serological typing were used as control. We conducted HLA-DR PCR-SSP typing on 18 samples from 6 true paternity trios, 16 samples from 8 true duos, 27 from 9 false trios, and 8 from 4 false duos. These DNAs from disputed paternity families had been previously tested for the parentage using polymerase chain reaction (PCR)-amplified short tandem repeat (STR) analysis.


RESULTS
No false positive nor false negative results were obtained in typing 28 positive control DNA samples from international standardized DNA Reference Panel for HLA Class II. Among the 20 DNA samples typed by microlymphocytotoxicity technique, the discordant typing results between HLA-DR PCR-SSP typing and serological typing were found in 3 (15%). In the family of true paternity, HLA-DR typing could not exclude any of alleged fathers and the pattern of inheritance was consistent with autosomal codominant. By HLA-DR typing alone, paternity in 2 alleged fathers out of 9 false trios and 2 alleged fathers out of 4 false duos could not be excluded.


CONCLUSIONS
Precise HLA-DR typing can be achieved by PCR-SSP analysis. Economic considerations preclude HLA-DR typing in routine parentage tests where STR typing is performed first.
Lin
Jeong-Shi
JS
Division of Transfusion Medicine, Department of Medicine, Taipei Veterans General Hospital, and National Yang-Ming University School of Medicine, Taipei, Taiwan, ROC. jslin@vghtpe.gov.tw
Tzeng
Cheng-Hwai
CH
Huang
De-Feng
DF
Lyou
Jau-Yi
JY
Chen
Ying-Ju
YJ
Hu
Hui-Yu
HY
Yung
Chien-Hui
CH
D000483:Alleles; D006684:HLA-DR Antigens / Q000235:genetics*; D006801:Humans; D008297:Male; D010334:Paternity; D016133:Polymerase Chain Reaction / Q000379:methods*; D020080:Tandem Repeat Sequences
D006684:HLA-DR Antigens