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Related Experiment Videos

KEL6 and KEL7 genotyping with sequence-specific primers.

Keli J Renoud1, Kathleen Barracchini, Karen M Byrne

  • 1Department of Transfusion Medicine, Warren G. Magnuson Clinical Center, National Institutes of Health, Bethesda, Maryland 20892-1184, USA.

Transfusion
|September 13, 2006
PubMed
Summary

A new genotyping assay using sequence-specific primer-polymerase chain reaction (SSP-PCR) can accurately determine Js(a) and Js(b) blood group alleles. Analysis of the KEL 1910-bp single-nucleotide polymorphism (SNP) is reliable for KEL6 and KEL7 genotyping.

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Area of Science:

  • Blood group genetics
  • Molecular diagnostics
  • Immunogenetics

Background:

  • Antibodies to Js(a) and Js(b) are clinically significant in transfusion medicine.
  • Reagent-quality anti-Js(a) and anti-Js(b) are not readily available.
  • A novel sequence-specific primer-polymerase chain reaction (SSP-PCR) assay was developed to genotype KEL6 and KEL7.

Purpose of the Study:

  • To evaluate the efficacy of an SSP-PCR genotyping assay for KEL6 and KEL7.
  • To compare genotyping results with Js(a) and Js(b) phenotypes.

Main Methods:

  • Developed four primer sets for selective amplification of KEL6 and KEL7.
  • Utilized two sets for the 1910 single-nucleotide polymorphism (SNP) and two for the 2019 SNP.
  • Compared SSP-PCR genotyping with Js(a) and Js(b) phenotyping in 64 subjects.

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Main Results:

  • SSP-PCRs demonstrated specificity for KEL6 and KEL7.
  • Genotyping at the 1910 SNP perfectly matched phenotyping results in all 64 subjects.
  • Genotyping at the 2019 SNP matched phenotyping in 49 subjects, with discrepancies attributed to an atypical KEL gene.

Conclusions:

  • The KEL 1910-bp SNP reliably genotypes KEL6 and KEL7.
  • The KEL 2019-bp SNP does not always correlate with Js phenotype due to an atypical KEL allele.
  • Analysis of the 1910-bp SNP is essential for accurate KEL6 and KEL7 genotyping.