PMID- 18954395
OWN - NLM
STAT- MEDLINE
DCOM- 20090203
LR  - 20090114
IS  - 1537-2995 (Electronic)
IS  - 0041-1132 (Linking)
VI  - 49
IP  - 1
DP  - 2009 Jan
TI  - Molecular typing of HLA genes using whole genome amplified DNA.
PG  - 57-63
LID - 10.1111/j.1537-2995.2008.01943.x [doi]
AB  - BACKGROUND: The outcome of clinical transplantation and a number of disease 
      susceptibilities show very strong associations with genetic variants within the 
      major histocompatibility complex, particularly in the human leukocyte antigen 
      (HLA) genes. A problem with many association studies is the lack of sufficient 
      DNA to perform multiple genetic analyses, particularly with transplantation 
      outcomes where donor and recipient DNA are often in short supply. This study 
      assesses whether a multiple-strand displacement whole genome amplification (WGA) 
      method could generate sufficient template of high quality to perform unbiased 
      amplification for analysis of the HLA-A, -B, -C, -DRB1, and -DQB1 genes. STUDY 
      DESIGN AND METHODS: A panel of DNA samples from various biological sources was 
      subjected to WGA reaction using Phi29 DNA polymerase. The HLA genotypes were 
      subsequently determined using standard polymerase chain reaction (PCR)-based 
      methods including sequence-specific oligonucleotide probes (PCR-SSOP, Luminex, 
      Luminex Corp.) and sequence-based typing (PCR-SBT). WGA products and original DNA 
      samples were used to determine the sensitivity of the Luminex assay; in addition, 
      reamplified WGA products were also genotyped. RESULTS: The WGA templates, as well 
      as serially amplified DNA for two successive rounds, yielded HLA genotypes fully 
      concordant with those determined for the original DNA samples. WGA products and 
      original DNA gave reproducible HLA-DQB1 genotypes with 100 to 10 ng of template. 
      Purification of the WGA products was required for successful PCR-SBT, but not for 
      the PCR-SSOP method. CONCLUSION: Our study suggests that WGA can be a reliable 
      method for generating unlimited DNA for medium- or high-resolution HLA typing 
      using the techniques described above.
FAU - Creary, Lisa E
AU  - Creary LE
AD  - Histocompatibility and Immunogenetics Research Group, Department of 
      Histocompatibility and Immunogenetics, Colindale Centre, NHSBT, NHSBT, London, 
      UK.
FAU - Girdlestone, John
AU  - Girdlestone J
FAU - Zamora, Jorge
AU  - Zamora J
FAU - Brown, Juliette
AU  - Brown J
FAU - Navarrete, Cristina V
AU  - Navarrete CV
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20081014
PL  - United States
TA  - Transfusion
JT  - Transfusion
JID - 0417360
SB  - IM
MH  - *Genome, Human
MH  - Genotype
MH  - Histocompatibility Testing/*methods
MH  - Humans
MH  - Polymerase Chain Reaction/*methods
MH  - Sensitivity and Specificity
EDAT- 2008/10/29 09:00
MHDA- 2009/02/04 09:00
CRDT- 2008/10/29 09:00
PHST- 2008/10/29 09:00 [pubmed]
PHST- 2009/02/04 09:00 [medline]
PHST- 2008/10/29 09:00 [entrez]
AID - TRF01943 [pii]
AID - 10.1111/j.1537-2995.2008.01943.x [doi]
PST - ppublish
SO  - Transfusion. 2009 Jan;49(1):57-63. doi: 10.1111/j.1537-2995.2008.01943.x. Epub 
      2008 Oct 14.