PMID- 25636098
OWN - NLM
STAT- MEDLINE
DCOM- 20150616
LR  - 20150131
IS  - 1003-9406 (Print)
IS  - 1003-9406 (Linking)
VI  - 32
IP  - 1
DP  - 2015 Feb
TI  - [Analysis of ambiguities in HLA sequencing-based typing and its solutions].
PG  - 44-8
LID - 10.3760/cma.j.issn.1003-9406.2015.01.010 [doi]
AB  - OBJECTIVE: To investigate the number and ratio of ambiguous allele combinations 
      from human leukocyte antigen (HLA) confirmatory test by sequencing based typing 
      for unrelated donor marrow transplantation, and to establish an efficient 
      strategy for identifying such ambiguities. METHODS: A total of 650 donor-receipt 
      samples were genotyped for 5 loci of the HLA gene using an Atria SBT commercial 
      kit. Exons 2, 3 and 4 of HLA-A, -B and -C, exon 2 of HLA-DRB1 and exons 2 and 3 
      of HLA-DQB1 were tested by routine HLA genotyping. The ratio of usual ambiguous 
      allele combination was calculated. The ambiguities were subjected to further 
      confirmatory test by PCR-SSP or PCR-SBT retest at outside of the routine 
      sequencing region. RESULTS: Among the 650 tested samples, the ratio of ambiguity 
      at HLA-A, B, C, DRB1 and DQB1 were 76.31% (496/650), 91.08% (592/650), 97.69% 
      (635/650), 88.62% (576/650) and 43.38% (141/650), respectively. A total of 36 
      ambiguous allele combinations inside the routine sequencing region and 22 
      ambiguous allele combinations outside of the routine sequencing region were 
      discovered. After removing rare alleles based on the Chinese common and well 
      documented (CWD) Allele Table (Version 1.01), 9 ambiguous CWD allele combinations 
      inside the routine sequencing region, including 3 located in HLA-B, HLA-C and 1 
      located in other three HLA loci were found. Ten ambiguous CWD allele combinations 
      outside of the routine sequencing region, including 4 located in HLA-C, -DRB1 and 
      1 in HLA-A, -B respectively were determined. All samples with ambiguous CWD 
      allele combinations could be distinguished by high-resolution PCR-SSP commercial 
      kits or PCR SBT retest at outside of the routine sequencing region. CONCLUSION: 
      The common and well documented allele combinations in sequencing-based typing at 
      five HLA loci have been analyzed. Our strategy may provide valuable information 
      for more efficient, low cost and accurate method for high resolution genotyping 
      of HLA genes.
FAU - Wang, Daming
AU  - Wang D
AD  - Immunogenetics Laboratory, Shenzhen Blood Center, Shenzhen, Guangdong 518035, P. 
      R. China. Email: zhihui_deng@sina.cn.
FAU - He, Liumei
AU  - He L
FAU - Zou, Hongyan
AU  - Zou H
FAU - Gao, Suqing
AU  - Gao S
FAU - Deng, Zhihui
AU  - Deng Z
LA  - chi
PT  - English Abstract
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Zhonghua Yi Xue Yi Chuan Xue Za Zhi
JT  - Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese 
      journal of medical genetics
JID - 9425197
RN  - 0 (HLA Antigens)
SB  - IM
MH  - Alleles
MH  - Genotype
MH  - HLA Antigens/*genetics
MH  - Humans
EDAT- 2015/01/31 06:00
MHDA- 2015/06/17 06:00
CRDT- 2015/01/31 06:00
PHST- 2015/01/31 06:00 [entrez]
PHST- 2015/01/31 06:00 [pubmed]
PHST- 2015/06/17 06:00 [medline]
AID - 940632010 [pii]
AID - 10.3760/cma.j.issn.1003-9406.2015.01.010 [doi]
PST - ppublish
SO  - Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2015 Feb;32(1):44-8. doi: 
      10.3760/cma.j.issn.1003-9406.2015.01.010.