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Error rate for HLA-B antigen assignment by serology: implications for proficiency testing and utilization of
M V Bozón1, J C Delgado, A Selvakumar
1Dana-Farber Cancer Institute, Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Tissue Antigens
|February 12, 1998
Summary
Serologic typing for HLA-B antigens shows significant errors. DNA-based methods like PCR-SSP and PCR-SSOP offer accurate HLA-B typing, improving clinical histocompatibility testing.
Area of Science:
- Immunogenetics
- Molecular Biology
- Histocompatibility Testing
Background:
- Historically, Human Leukocyte Antigen (HLA) class I typing relied on serological methods.
- DNA-based typing strategies are increasingly adopted to enhance accuracy and address limitations of serology, particularly for diverse populations.
- Serological methods can be expensive and prone to inaccuracies, impacting clinical histocompatibility testing.
Purpose of the Study:
- To compare the accuracy of serological HLA-B typing with two DNA-based methods: PCR-SSP and PCR-SSOP.
- To evaluate the effectiveness of combining serological and DNA-based typing for improved HLA-B antigen resolution.
- To assess the utility of DNA-based methods in improving clinical histocompatibility and proficiency testing.
Main Methods:
- A double-blind comparison of serology, PCR-SSP, and PCR-SSOP was conducted on 40 samples for HLA-B specificities.
- A second panel of 20 donor samples with ambiguous serological results was analyzed using PCR-SSP and PCR-SSOP.
- Polymerase chain reaction amplification with sequence-specific primers (PCR-SSP) and PCR amplification and subsequent hybridization with sequence-specific oligonucleotide probes (PCR-SSOP) were employed.
Main Results:
- Serological typing resulted in 22.5% misassignments of HLA-B antigens.
- Complete concordance was observed between the two PCR-based typing methods (PCR-SSP and PCR-SSOP).
- Both PCR methods accurately identified HLA-B antigens in samples with incomplete or ambiguous serological results.
Conclusions:
- DNA-based typing techniques, such as PCR-SSP and PCR-SSOP, provide more accurate HLA-B typing results compared to serology alone.
- Combining serological testing with limited DNA-based typing achieves a resolution of HLA-B antigen assignment equivalent to WHO-HLA Committee definitions.
- Serological reagents alone are insufficient for achieving the required resolution in routine clinical histocompatibility and proficiency testing.