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Related Experiment Videos

Rapid HLA-DRB1 genotyping by nested PCR amplification.

G Bein1, R Gläser, H Kirchner

  • 1Institute of Immunology and Transfusion Medicine, University of Lübeck, Medical School, FRG.

Tissue Antigens
|February 1, 1992
PubMed
Summary

This study presents a rapid DNA typing method for human leukocyte antigen (HLA) class II alleles using nested polymerase chain reaction (PCR). This faster method is suitable for typing organ donors for HLA-DR matching in transplantation.

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Tissue antigens·2010

Area of Science:

  • Immunogenetics
  • Molecular Biology
  • Transplantation Science

Background:

  • Current human leukocyte antigen (HLA) class II genotyping methods are time-consuming, exceeding one working day.
  • Existing methods are unsuitable for rapid typing of cadaveric organ donors.

Purpose of the Study:

  • To develop a rapid DNA typing method for HLA class II alleles.
  • To enable timely HLA-DR matching for organ transplantation.

Main Methods:

  • Designed specific oligonucleotide primer sets for nested polymerase chain reaction (PCR) amplification.
  • Amplified DRB genes exon 2 and DRw52-group-specific part of DRB1 exon 2 directly from cell lysates.
  • Utilized 18 nested allele- or group-specific primer pairs in the second amplification round.

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Main Results:

  • Successfully typed all serological HLA-DR specificities (DR1-DRw18) by detecting amplified DNA after agarose gel electrophoresis.
  • Demonstrated complete refractoriness to amplification with mismatched terminal 3'-nucleotide bases.
  • Completed the entire assay in less than one working day.

Conclusions:

  • Developed a rapid, easy-to-perform DNA typing assay for HLA-DR alleles.
  • This method is suitable for prospective HLA-DR matching of organ donors in renal transplantation.