- Author:
Suqing GAO
1
;
Daming WANG
;
Yunping XU
;
Zhihui DENG
Author Information
- Publication Type:Journal Article
- MeSH: Alleles; DNA Primers; genetics; Exons; Genotype; HLA-DQ beta-Chains; genetics; Histocompatibility Testing; methods; Humans; Polymerase Chain Reaction; methods
- From: Chinese Journal of Medical Genetics 2014;31(4):496-498
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the reason for HLA-DQB1 allele dropout during routine sequence-based typing(SBT) in order to improve the accuracy of typing.
METHODSTwo thousand samples derived from HLA high-resolution typing laboratory were typed for HLA-DQB1 locus using an AlleleSEQR HLA-DQB1 SBT kit. Non-conclusive results and "abnormal" sequencing samples were retyped using a LABType rSSO HD HLA-DQB1 kit and further analyzed with both sequence-specific primers and group-specific primers and sequenced for haplotype analysis.
RESULTSAmong the 2000 samples, 2 samples with no conclusive result were identified. The heterozygosity was confirmed with both the LAB Type SSO HD HLA-DQB1 kit and PCR-SBT in house method. Subsequent HLA-DQB1 cloning and haplotype sequencing have elucidated that HLA-DQB1*02:02 dropped out at exon 2 for the first sample and HLA-DQB1*02:01:01 dropped out at exon 2 for the second sample during PCR amplification. No novel nucleotide mutation was found.
CONCLUSIONOur results indicated that preferential amplification at exon 2 of DQB1 may result in allele dropout in exon 2 sequences during HLA-DQB1 SBT test. This may provide useful information for HLA genotyping.