Title: Use of dried blood spots for the determination of genetic variation of IL-10, Killer Immunoglobulin-like Receptor and HLA Class I genes
Authors: Ndlovu BG, Danaviah S, Moodley E, Ghebremichael M, Bland R, Viljoen J, Newell M-L, Ndung'u T, and Carr WH.
Journal: Tissue Antigens,79(2):114-22 (2011)
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 sequencespecific 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.
Download: Full text paper
Citation: Ndlovu BG, Danaviah S, Moodley E, Ghebremichael M, Bland R, Viljoen J, Newell M-L, Ndung'u T, and Carr WH. Use of dried blood spots for the determination of genetic variation of IL-10, Killer Immunoglobulin-like Receptor and HLA Class I genes Tissue Antigens,79(2):114-22 (2011).