南方汉族强直性脊柱炎患者及健康人群HLA-B27基因的分子多态性及分布

Molecular polymorphism and the distribution of HLA B-27 subtypes in southern Chinese Han patients with Ankylosing Spondylitis and healthy controls

目的研究南方汉族强直性脊柱炎(AS)患者及健康人群HLA-B27亚型等位基因的多态性和分布特点。方法采用HLA测序分型方法,对收集到的46份B*27阳性的南方汉族AS患者血样以及随机选择的80份经rS-SOHLA流式磁珠低分辨检测为B*27阳性的造血干细胞南方汉族捐献者血样,做HLA-B基因的第2—4外显子测序分型,测序反应产物用ABI PrismTM3730测序仪检测,Assign3.5分析软件分析结果。对检出的模棱两可结果及"罕见型"等位基因,辅以PCR-SSP法HLA-B27高分辨基因分型。结果46名南方汉族AS患者中,共检出4个HLA-B*27相关的等位基因:B*2704的检出比例最高,为82.98%(39/47);其次是B*2705,为12.77%(6/47);B*2707和B*2724各为2.13%(1/47)。80名南方汉族造血干细胞捐献者中,共检出7种B*27相关等位基因:以B*2704的检出比例最高,为57.32%(47/82);其次为是B*2705,为26.83%(22/82);B*2706和B*2707各为6.10%(5/82);B*2703、B*2715和B*2724各为1.22%(1/82)。结论南方汉族AS患者HLA-B*27基因中以B*2704最为常见,健康人群以B*2704和B*2705等位基因为主要亚型。

Objective To study the molecular polymorphism and the distribution of HLA-B ＊ 27 subtypes in southern Chinese Han patients with Ankylosing Spondylitis and healthy controls. Methods A total of 46 samples form southern Chi- nese Han patients with AS and 80 non-related blood samples from healthy peripheral blood stem cell donors with B＊ 27-pos- itive identified by rSSO Lumminex flow array assay were subjected to sequencing analysis of exon 2 , 3 and 4 of HLA-B gene by the sequence-based typing, the purified products of sequencing reaction were electrophoresed on ABI 3730 DNA sequen- cer and the designation of HLA-B＊ 27 allele was accomplished using the Assign3.5 software. The ambiguities and the detec- ted ＂rare＂ alleles were confirmed using the PCR-SSP commercial kit. Results In the 46 B＊27-positive patients with the diagnosis of AS, four alleles, namely B＊ 2704, B＊ 2705, B＊ 2707 and B＊ 2724 were determined. The frequencies for these four alleles were 82.98% （39/47）, 12.77% （6/47）, 2.13% （1/47） and 2.13% （ 1/47）, respectively. In the 80 B＊ 27- positive control individuals, seven B＊ 27 related alleles were identified. The frequency for the two dominant subtype B＊ 2704 and B＊ 2705 were 57.32% （47/82） and 26.83% （22/82）, respectively. Both the B＊ 2706 and B＊ 2707 were ob- served 5 times with a frequency of 6.10% （5/82）, three alleles B＊2703, B＊2715 and B＊2724 were detected only once with a frequency of 1.22% （1/82）. Conclusion Our study shows that HLA-B＊ 2704 and B＊ 2705 were the predominant subtypes in normal healthy controls, however, B＊ 2704 was the predominant subtype for the AS group in southern Chinese Han patients.