[DNA typing for HLA-DR in donor-recipients of cadaveric transplantation].
OBJECTIVE
To accurately allocate donor-recipients of HLA-matching and improve long-term graft survival, genotyping method for HLA-DR alleles by polymerase chain reaction with sequence-specific primers (PCR-SSP) was established and applied to renal transplantation.


METHODS
Thirty primers were designed and synthesized according to the HLA-DR nucleotide sequences. Genomic DNAs were prepared by a rapid salting-out method. A rapid genotyping method of PCR-SSP was set up by PCR technique and applied to HLA-DR typing in 14 cell lines DNA and 171 individuals of donor-recipients of cadaveric transplantation. The amplification was accomplished by 34 cycles consisting of denaturation at 94 degrees C for 30 seconds, annealing at 60 degrees C for 50 seconds, and extension at 72 degrees C for 40 seconds. The specificity of matching was determined against a panel of standard DNA, analysis with restriction endonucleases and Southern hybridization.


RESULTS
HLA-DR alleles of all 171 samples and 14 cell line DNAs were able to be typed by PCR-SSP. The size of specific products was consistent with the size of calculation. The overall time of genotyping was only 5 hours. No false positive or false negative typing results were discovered. The typing results were confirmed by analysis with endonucleases and hybridization. The specificity and reproducibility were 100%.


CONCLUSIONS
Genotyping for HLA-DR by PCR-SSP is a rapid and accurate matching technique, suitable for organ transplantation, especially allocation of donor-recipients of cadaveric transplantation.
Tan
J
J
Department of Urology, Shanghai First People's Hospital.
Xie
T
T
Xu
Q
Q
D000483:Alleles; D004247:DNA / Q000145:classification; D005838:Genotype; D006085:Graft Survival; D006684:HLA-DR Antigens / Q000145:classification / Q000235:genetics*; D006650:Histocompatibility Testing; D006801:Humans; D016030:Kidney Transplantation; D016133:Polymerase Chain Reaction; D012189:Retrospective Studies; D014019:Tissue Donors
D006684:HLA-DR Antigens; D004247:DNA