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我们用聚合酶链式反应(PCR)和序列特异性寡核苷酸(SSO)探针技术检测了IgAN患者HLA-DRB1、DQA1、DQB1等位基因频率,并探讨HLA与IgAN之间的关联及其临床意义。 一、材料与方法 1.病例来源:随机选择经肾穿刺免疫荧光证实的IgAN患者47例,其中男性24人,女性23人,平均年龄32岁。临床排除SLE,过敏性紫癜,慢性肝脏病等继发性IgAN。对照组:数据由上海第二医科大学免疫研究所分子遗传组提供,共包括92例健康者。 2.方法:提取外周血白细胞基因组DNA,利用PCR技术扩增HLA-Ⅱ类基因的第二外显子,末端标记SSO探针进行斑点杂交,按Tm值调整洗膜后-70℃
We examined the allele frequencies of HLA-DRB1, DQA1 and DQB1 in patients with IgAN using polymerase chain reaction (PCR) and sequence-specific oligonucleotide (SSO) probe techniques and explored the association between HLA and IgAN and Its clinical significance. Materials and Methods 1. Case Source: Randomly selected 47 cases of IgAN confirmed by renal biopsy, including 24 males and 23 females, with an average age of 32 years. Clinical excluded SLE, Henoch-Schonlein purpura, chronic liver disease secondary IgAN. Control group: The data provided by Shanghai Jiao Tong University Institute of Immunology molecular genetic group, a total of 92 healthy subjects. Methods: The genomic DNA of peripheral blood leukocytes was extracted. The second exon of HLA-Ⅱ gene was amplified by PCR, and terminal SSO probe was used for spot blot hybridization. The Tm value was adjusted to -70 ℃