title: Use of dried blood spots for the determination of genetic variation of interleukin-10, killer immunoglobulin-like receptor and HLA class I genes.
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.
authors: HIV Pathogenesis Programme, Doris Duke Medical Research Institute, Nelson R. Mandela School of Medicine, University of KwaZulu-Natal, Durban, South Africa.
authors: B G
authors: BG
authors: Ndlovu
authors: S
authors: S
authors: Danaviah
authors: E
authors: E
authors: Moodley
authors: M
authors: M
authors: Ghebremichael
authors: R
authors: R
authors: Bland
authors: J
authors: J
authors: Viljoen
authors: M-L
authors: ML
authors: Newell
authors: T
authors: T
authors: Ndung'u
authors: W H
authors: WH
authors: Carr
mesh_terms: D000293:Adolescent; D000328:Adult; D004247:DNA / Q000032:analysis / Q000235:genetics; D016172:DNA Fingerprinting / Q000379:methods*; D059788:Dried Blood Spot Testing; D005260:Female; D014644:Genetic Variation; D005838:Genotype; D015395:Histocompatibility Antigens Class I / Q000235:genetics* / Q000276:immunology; D006801:Humans; D016753:Interleukin-10 / Q000235:genetics* / Q000276:immunology; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D020033:Protein Isoforms / Q000235:genetics* / Q000276:immunology; D054340:Receptors, KIR / Q000235:genetics* / Q000276:immunology; D012680:Sensitivity and Specificity
chemical_list: D015395:Histocompatibility Antigens Class I; D020033:Protein Isoforms; D054340:Receptors, KIR; D016753:Interleukin-10; D004247:DNA