Objectives: Human leukocyte antigen (HLA) B27 is a susceptibility allele of ankylosing spondylitis (AS), and HLA-B27 antigen typing is an important indicator for clinical diagnosis of AS, but current typing methods such as sequence specific primer polymerase chain reaction (PCR-SSP) still possess limitation. Therefore, this study aims to analyze the correlation between B27 subtypes and susceptibility to AS in Hunan Province by applying high-resolution polymerase chain reaction-sequence-based typing (PCR-SBT).
Methods: Peripheral blood of 116 patients with suspected AS (suspected AS group) and 121 healthy volunteers (control group) admitted to the Second Xiangya Hospital from January 2020 to December 2020 were collected for HLA-B genotyping by PCR-SBT. Among the patients in the suspected AS group, 23 patients were finally diagnosed with AS (confirmed AS group), and the remaining 93 undiagnosed patients served as the non-confirmed AS group. PCR-SBT and PCR-SSP were used to detect HLA-B27 typing in 116 patients with suspected AS, and the results of the 2 methods were compared.
Results: The HLA-B27 allele frequency in the suspected AS group was significantly higher than that in the control group [11.63% vs 2.48%; P<0.001, odds ratio (OR)=5.18, 95% confidence interval (CI) 2.097 to 12.795]. B*27:04, B*27:05, B*27:06, and B*27:07 were detected in the suspected AS group and the control group. The frequency of the B*27:04 allele in the suspected AS group was significantly higher than that in the control group (9.48% vs 1.24%; P<0.001, OR=8.346, 95% CI 2.463 to 28.282). The positive rate of B27 in the suspected AS group and the confirmed AS group (B27+/+ and B27+/-) was significantly higher than that in the control group (χ2=16.579, P<0.001; χ2=94.582, P<0.001, respectively). Among the confirmed AS group, 21 were HLA-B27 carriers, and the B27 positive rate in the confirmed AS group was 91.3%. PCR-SBT could achieve high resolution typing of the HLA-B gene locus, with higher sensitivity, specificity, positive predictive value, negative predictive value, and accuracy than PCR-SSP.
Conclusions: PCR-SBT typing analysis shows a strong correlation between HLA-B * 27:04 and AS in Hunan province. The PCR-SBT method can be used as the preferred option for the auxiliary diagnosis of clinical AS.
目的: 人类白细胞抗原(human leukocyte antigen，HLA)B27是强直性脊柱炎(ankylosing spondylitis，AS)的易感等位基因，HLA-B27抗原分型是临床诊断AS的重要指标，但目前的分型方法如序列特异引物聚合酶链反应技术(polymerase chain reaction-sequence specific primer，PCR-SSP)等仍存在一定的缺陷。因此，本研究旨在应用高分辨率聚合酶链反应直接测序分型法(polymerase chain reaction-sequence-based typing，PCR-SBT)分析B27亚型与湖南地区强直性脊柱炎易感性的相关性。方法: 收集2020年1至12月在湘雅二医院门诊就医的116例疑似AS患者(疑似AS组)和121名健康志愿者(对照组)的外周血，采用PCR-SBT进行HLA-B基因分型。在疑似AS组患者中，23名患者最终被确诊为AS(AS确诊组)，其余的93名未能确诊的患者为非AS确诊组。采用PCR-SBT和PCR-SSP检测116例疑似AS患者的HLA-B27分型，并比较2种方法的检测结果。结果: 疑似AS组HLA-B27等位基因频率显著高于对照组[11.63% vs 2.48%；P<0.001，优势比(odds ratio，OR)=5.18，95%置信区间(confidence interval，CI) 2.097~12.795]。疑似AS组和对照组均检出B*27:04、B*27:05、B*27:06、B*27:07。疑似AS组B*27:04等位基因频率明显高于对照组(9.48% vs 1.24%；P<0.001，OR=8.346，95% CI 2.463~28.282)。疑似AS组和AS确诊组B27阳性率(B27+/+和B27+/-)均显著高于对照组(分别χ2=16.579，P<0.001；χ2=94.582，P<0.001)。在AS确诊组中，21例为HLA-B27携带者，AS确诊组B27阳性率为91.3%。PCR-SBT方法可对HLA-B基因位点进行高分辨分型，其敏感度、特异度、阳性预测值、阴性预测值、准确度均高于PCR-SSP法。结论: 应用PCR-SBT检测出HLA-B*27:04与湖南地区AS具有强相关性。PCR-SBT可作为临床AS辅助诊断的优选方案。.
目的: 人类白细胞抗原(human leukocyte antigen，HLA)B27是强直性脊柱炎(ankylosing spondylitis，AS)的易感等位基因，HLA-B27抗原分型是临床诊断AS的重要指标，但目前的分型方法如序列特异引物聚合酶链反应技术(polymerase chain reaction-sequence specific primer，PCR-SSP)等仍存在一定的缺陷。因此，本研究旨在应用高分辨率聚合酶链反应直接测序分型法(polymerase chain reaction-sequence-based typing，PCR-SBT)分析B27亚型与湖南地区强直性脊柱炎易感性的相关性。
方法: 收集2020年1至12月在湘雅二医院门诊就医的116例疑似AS患者(疑似AS组)和121名健康志愿者(对照组)的外周血，采用PCR-SBT进行HLA-B基因分型。在疑似AS组患者中，23名患者最终被确诊为AS(AS确诊组)，其余的93名未能确诊的患者为非AS确诊组。采用PCR-SBT和PCR-SSP检测116例疑似AS患者的HLA-B27分型，并比较2种方法的检测结果。
结果: 疑似AS组HLA-B27等位基因频率显著高于对照组[11.63% vs 2.48%；P<0.001，优势比(odds ratio，OR)=5.18，95%置信区间(confidence interval，CI) 2.097~12.795]。疑似AS组和对照组均检出B*27:04、B*27:05、B*27:06、B*27:07。疑似AS组B*27:04等位基因频率明显高于对照组(9.48% vs 1.24%；P<0.001，OR=8.346，95% CI 2.463~28.282)。疑似AS组和AS确诊组B27阳性率(B27+/+和B27+/-)均显著高于对照组(分别χ 2=16.579，P<0.001；χ 2=94.582，P<0.001)。在AS确诊组中，21例为HLA-B27携带者，AS确诊组B27阳性率为91.3%。PCR-SBT方法可对HLA-B基因位点进行高分辨分型，其敏感度、特异度、阳性预测值、阴性预测值、准确度均高于PCR-SSP法。
结论: 应用PCR-SBT检测出HLA-B*27:04与湖南地区AS具有强相关性。PCR-SBT可作为临床AS辅助诊断的优选方案。
Keywords: ankylosing spondylitis; genetic susceptibility; human leukocyte antigen-B*27:04; polymerase chain reaction sequence-based typing.