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High density FTA plates serve as efficient long-term sample storage for HLA genotyping.

V Lange1, K Arndt, C Schwarzelt

  • 1DKMS Life Science Lab, Dresden, Germany.

Tissue Antigens
|January 9, 2014
PubMed
Summary

Dried blood spots (DBS) on FTA plates are suitable for high-resolution human leukocyte antigen (HLA) sequencing. Proper DNA extraction methods ensure reliable results from long-term stored DBS and whole blood samples.

Keywords:
DNA extractionFTA platesdried blood spotsgenotypinghuman leukocyte antigensequencing-based typing

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Area of Science:

  • Genomics
  • Immunogenetics
  • Molecular Biology

Background:

  • Dried blood spots (DBS) offer a stable alternative to frozen storage for biological samples.
  • The suitability of DBS for high-resolution human leukocyte antigen (HLA) allele sequencing remains debated.
  • Long-term storage of biological samples impacts DNA quality and downstream applications.

Purpose of the Study:

  • To evaluate the efficacy of DNA extraction from DBS stored on FTA plates for high-resolution HLA sequencing.
  • To identify optimal DNA extraction methods for maximizing DNA yield and quality from stored DBS.
  • To determine if frozen whole blood can be transferred to filter paper without compromising HLA genotyping.

Main Methods:

  • DNA extraction from DBS stored for up to four years using six distinct methods.
  • High-resolution sequencing of human leukocyte antigen (HLA) alleles.
  • Comparison of DNA quality and quantity across different extraction protocols.
  • Assessment of HLA genotyping accuracy from long-term stored samples.

Main Results:

  • Specific DNA extraction protocols were identified that yield sufficient high-quality DNA from DBS for reliable high-resolution HLA sequencing.
  • Frozen whole blood samples stored for several years maintained their suitability for HLA genotyping after transfer to filter paper.
  • The storage duration of DBS on FTA plates did not preclude successful high-resolution HLA sequencing when appropriate methods were used.

Conclusions:

  • DNA derived from DBS stored on high-density FTA plates is suitable for high-resolution HLA sequencing.
  • The selection of appropriate DNA extraction protocols is critical for successful genotyping from stored DBS.
  • Transferring long-term stored frozen whole blood to filter paper does not compromise HLA genotyping accuracy.