Zhang Bo, Liu Limin, Zhou Tian, Shi Xiaoli, Wu Haijing, Xiang Zhongyuan, Zhao Ming, Lu Qianjin
Department of Dermatology, Second Xiangya Hospital, Central South University, Hunan Key Laboratory of Medical Epigenomics, Changsha, Hunan, China; Research Unit of Key Technologies of Diagnosis and Treatment for Immune-related Skin Diseases, Chinese Academy of Medical Sciences (2019RU027), Changsha, Hunan, China.
Department of Clinical Laboratory, Second Xiangya Hospital, Central South University, Changsha, Hunan, China.
Clin Immunol. 2020 Dec;221:108612. doi: 10.1016/j.clim.2020.108612. Epub 2020 Oct 16.
Systemic lupus erythematosus (SLE) is a complex heterogenous autoimmune disease that can be challenging to diagnose. We previously identified the IFN-induced protein 44-like (IFI44L) methylation marker for SLE diagnosis, which can be detected by pyrosequencing. Although the previous technique has high sensitivity and specificity, it requires special equipment and high cost for detection. Here, we established a high-resolution melting-quantitative polymerase chain reaction (HRM-qPCR) assay to detect the methylation of IFI44L promoter for the diagnosis of SLE. The result was determined according to the standard melting curve of the methylation level of the IFI44L promoter region. The sensitivity was 88.571% and the specificity was 97.087%. The HRM-qPCR and pyrosequencing results presented good consistency when both methods were used to detect the methylation of the IFI44L promoter for SLE diagnosis. Furthermore, the HRM-qPCR method can be used to distinguish SLE from other autoimmune diseases, infectious diseases and virus-related cancers.
系统性红斑狼疮（SLE）是一种复杂的异质性自身免疫性疾病，诊断具有挑战性。我们之前鉴定了用于SLE诊断的干扰素诱导蛋白44样（IFI44L）甲基化标志物，可通过焦磷酸测序检测。尽管先前的技术具有高灵敏度和特异性，但它需要特殊设备且检测成本高。在此，我们建立了一种高分辨率熔解定量聚合酶链反应（HRM-qPCR）检测方法来检测IFI44L启动子的甲基化以诊断SLE。根据IFI44L启动子区域甲基化水平的标准熔解曲线确定结果。灵敏度为88.571%，特异性为97.087%。当两种方法都用于检测IFI44L启动子的甲基化以诊断SLE时，HRM-qPCR和焦磷酸测序结果呈现出良好的一致性。此外，HRM-qPCR方法可用于区分SLE与其他自身免疫性疾病、传染病和病毒相关癌症。
