Use of Dried Blood Spots for the Determination of Genetic Variation of interleukin-10, Killer Immunoglobulin-Like Receptor and HLA Class I Genes

Tissue Antigens. 2012 Feb;79(2):114-22. doi: 10.1111/j.1399-0039.2011.01807.x. Epub 2011 Nov 22.

Abstract

Optimal methods for using dried blood spots (DBSs) for population genetics-based studies have not been well established. Using DBS stored for 8 years from 21 pregnant South African women, we evaluated three methods of gDNA extraction with and without whole-genome amplification (WGA) to characterize immune-related genes: interleukin-10 (IL-10), killer immunoglobulin-like receptors (KIRs) and human leukocyte antigen (HLA) class I. We found that the QIAamp DNA mini kit yielded the highest gDNA quality (P< 0.05; Wilcoxon signed rank test) with sufficient yield for subsequent analyses. In contrast, we found that WGA was not reliable for sequence-specific primer polymerase chain reaction (SSP-PCR) analysis of KIR2DL1, KIR2DS1, KIR2DL5 and KIR2DL3 or high-resolution HLA genotyping using a sequence-based approach. We speculate that unequal template amplification by WGA underrepresents gene repertoires determined by sequence-based approaches.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • DNA / analysis
  • DNA / genetics
  • DNA Fingerprinting / methods*
  • Dried Blood Spot Testing*
  • Female
  • Genetic Variation
  • Genotype
  • Histocompatibility Antigens Class I / genetics*
  • Histocompatibility Antigens Class I / immunology
  • Humans
  • Interleukin-10 / genetics*
  • Interleukin-10 / immunology
  • Middle Aged
  • Polymerase Chain Reaction
  • Pregnancy
  • Protein Isoforms / genetics*
  • Protein Isoforms / immunology
  • Receptors, KIR / genetics*
  • Receptors, KIR / immunology
  • Sensitivity and Specificity

Substances

  • Histocompatibility Antigens Class I
  • Protein Isoforms
  • Receptors, KIR
  • Interleukin-10
  • DNA