Open AccessUse of dried blood spots for the determination of genetic variation of interleukin‐10, killer immunoglobulin‐like receptor and HLA class I genes
Author(s) -
Ndlovu B. G.,
Danaviah S.,
Moodley E.,
Ghebremichael M.,
Bland R.,
Viljoen J.,
Newell M.L.,
Ndung'u T.,
Carr W. H.
Publication year - 2012
Publication title -
tissue antigens
Language(s) - English
Resource type - Journals
eISSN - 1399-0039
pISSN - 0001-2815
DOI - 10.1111/j.1399-0039.2011.01807.x
Subject(s) - biology , genotyping , human leukocyte antigen , genomic dna , polymerase chain reaction , genetics , primer (cosmetics) , microbiology and biotechnology , gene , genotype , population , antigen , medicine , chemistry , environmental health , organic chemistry
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.