J Clin Pathol Qual Control.
1997 Jun;19(1):215-231.
A Comparative Study of HLA-DR Typing by Polymerase Chain Reaction with Sequence-Specific Primers and Serology
- Affiliations
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- 1Department of Clinical Pathology, College of Medicine, Keimyung University, Taegu, Korea.
- 2Department of Clinical Pathology, Masan-Samsung Hospital, Masan, Korea.
Abstract
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BACKGROUND: Matching for the HLA-A, B, and more importantly, DR between donor and recipient has a strong influence on long-term graft survival rates in organ transplantation, especially kidney and bone marrow. Serologic typing has long been used, but due to its high error rates of up to 25%, there is a clinical need for exact DNA typing method. Recently polymerase chain reaction (PCR)-based DNA typing techniques have been successfully used, among which PCR with sequence specific primer (PCR-SSP) is known to be reliable and by its simple post-amplification step, sufficiently rapid for HLA-DR typing in emergency situation such as cadaveric transplantation.
METHODS
In this study, to evaluate the relative performance and reliability of the PCR-SSP method compared to serological typing, total 100 clinical samples from 85 renal transplant patients and 15 patients with other diseases were assayed for the HLA-DR typing by both serology and PCR-SSP. According to Wetzsteon et al., confidence level assignments of serological typing were made: c1A and c1B, no problem in assignment; c1C and c1D, difficult in assignment. Fifteen B-lymphoblastoid cell lines were used as a source of reference DNA to confirm the specificity of PCR-SSP.
RESULTS
All the amplification patterns using cell lines were easy to interpret and identical to the expected patterns. In serological typing using clinical samples, 60.0% were assigned as c1A and c1B and 40.0% were assigned as c1C and c1D, and there were 29 (29.0%) discrepancies in HLA-DR typing between serology and PCR-SSP. Of these, four (13.8%) were serological failure; 18 (62.0%) assignments of other alleles by PCR-SSP to serological blank; four (13.8%) differently assigned by serology and PCR-SSP; three (10.3%) allele splits not assignable by serology. But there was no single case of PCR-SSP failure.
CONCLUSIONS
HLA-DR typing by the PCR-SSP technique, especially a rapid modified method of PCR-SSP, is ideally suited for analyzing small number of samples simultaneously and is an alternative to serological typing in routine clinical practice including donor-recipient matching in cadaveric transplantations.
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