Use of dried blood spots for the determination of genetic variation of interleukin-10, killer immunoglobulin-like receptor and HLA class I genes

Ndlovu, B.G., Danaviah, S., Moodley, E., Ghebremichael, M., Bland, R., Viljoen, J., Newell, M.-L., Ndung'u, T. and Carr, W.H. (2011) 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, 79(2), pp. 114-122. (doi: 10.1111/j.1399-0039.2011.01807.x)

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.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Bland, Dr Ruth
• Authors: Ndlovu, B.G., Danaviah, S., Moodley, E., Ghebremichael, M., Bland, R., Viljoen, J., Newell, M.-L., Ndung'u, T., and Carr, W.H.
• College/School: College of Medical Veterinary and Life Sciences > School of Medicine, Dentistry & Nursing > Clinical Specialities
• Journal Name: Tissue Antigens
• ISSN: 0001-2815