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HLA class II genotyping: two assay systems compared
J Thonnard1, F Deldime, M Heusterspreute
1Clinical Laboratory of Molecular Biology, Louvain University Medical School, Cliniques Universitaires St-Luc, Brussels, Belgium.
Clinical Chemistry
|April 1, 1995
Summary
Comparing DNA-based human leukocyte antigen (HLA) typing methods, this study found both PCR-based techniques reliable for routine HLA-DRB typing. Combining methods enhances accuracy for high-resolution allelic typing.
Area of Science:
- Immunogenetics
- Molecular Biology
- Clinical Laboratory Science
Background:
- Human leukocyte antigen (HLA) gene polymorphism presents challenges for clinical laboratories in selecting optimal DNA-based typing strategies.
- The extreme polymorphism necessitates reliable and efficient HLA typing methods for various clinical applications.
Purpose of the Study:
- To compare the accuracy and efficiency of two DNA-based methods for human leukocyte antigen - DRB (HLA-DRB) typing: Inno-LiPA and PCR-restriction fragment length polymorphism (RFLP).
- To evaluate the suitability of each method for routine large-scale typing versus high-resolution allelic typing.
Main Methods:
- Utilized polymerase chain reaction (PCR) for amplification of HLA-DRB genes.
- Employed Inno-LiPA (PCR followed by reverse dot-blot hybridization) and PCR-RFLP for typing analysis.
Main Results:
- Both Inno-LiPA and PCR-RFLP demonstrated high concordance (97%) and reliability for HLA-DRB typing.
- The Inno-LiPA assay proved convenient for large-scale routine typing.
- Neither method alone achieved sufficient accuracy for high-resolution allelic typing, but their combined use improved accuracy.
Conclusions:
- Both evaluated DNA-based methods are reliable for HLA-DRB typing, with Inno-LiPA being efficient for routine use.
- Combining Inno-LiPA and PCR-RFLP enhances the accuracy of high-resolution allelic typing, addressing limitations of individual methods.
