Autoantibodies undetectable by chemiluminescent enzyme immunoassay require extended antigen-antibody reaction time for detection
Autoantibodies undetectable by chemiluminescent enzyme immunoassay require extended antigen-antibody reaction time for detection
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化学发光酶联免疫分析无法检测到的自身抗体需要延长抗原抗体反应时间才能检测
DOI：
10.1111/bjd.17121
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Shimizu H
中科院分区：
文献类型：
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作者：
Mai Y;Ujiie H;Higashi T;Yamagami J;Iwata H;Shimizu H
寻常型天疱疮（Pemphigus vulgaris， PV）是一种影响粘膜和皮肤的自身免疫性起泡疾病。PV产生针对桥粒黏附分子桥粒蛋白（Dsg） 3和Dsg1的自身抗体。为了检测这些自身抗体，酶联免疫吸附试验（ELISA）已被广泛使用。近年来，化学发光酶免疫分析法（CLEIA）得到了发展。CLEIA是一种使用化学发光技术的自动化系统，比ELISA具有更宽的动态范围和更短的检测时间。因此，CLEIA已经开始取代ELISA检测自身抗体。然而，最近的报告显示，一些天疱疮病例具有独特的抗dsg3或抗dsg1自身抗体，CLEIA无法检测到，但ELISA可检测到。4,5为了解决这一差异，我们分析了CLEIA检测不到抗dsg3或抗dsg1自身抗体的6例PV患者的血清。患者特征如图1 （a, b）所示。本研究依据《赫尔辛基宣言》原则进行，所有参与者均给予书面知情同意。病例2的临床资料我们之前已经报道过。病例3、5和6的血清曾用于Fujio等人的研究。4所有病例均被诊断为PV。病例1在口腔糜烂复发时采集血清，病例5在口腔糜烂缓解期采集血清。其余病例均为PV活跃期。所有血清在CLEIA中均未与Dsg3或Dsg1发生反应（STACIA MEBLux测试Dsg3和Dsg1； MBL，东京，日本）。1 . a, b)。然而，通过ELISA检测抗Dsg1或抗Dsg3自身抗体（mes针孔-2粘连蛋白试验Dsg3和Dsg1； MBL）（图2）。1a, b)，其中在CLEIA和ELISA中使用相同的Dsg3和Dsg1重组蛋白。所有病例均有IgG自身抗体对正常人表皮细胞间隙产生反应（数据未显示）。这些结果证实这些血清含有抗dsg3或抗dsg1自身抗体，ELISA检测不到，而CLEIA检测不到。因此，为了解决这些检测结果之间的差异，我们重点研究了CLEIA之间抗原抗体反应的孵育时间差异很大
DEAR EDITOR, Pemphigus vulgaris (PV) is an autoimmune blistering disease affecting the mucous membranes and the skin. 1 PV produces autoantibodies targeting the desmosomal adhesion molecules desmoglein (Dsg) 3 and Dsg1. 2 To detect these autoantibodies, enzyme-linked immunosorbent assay (ELISA) has been widely used. Recently, chemiluminescent enzyme immunoassay (CLEIA) has been developed. 3 CLEIA is an automated system that uses chemiluminescent technology with a wider dynamic range and a shorter assay time than those of ELISA. 3 Therefore, CLEIA has started to replace ELISA for autoantibody detection. However, recent reports have shown a few cases of pemphigus with unique anti-Dsg3 or anti-Dsg1 autoantibodies that were undetectable by CLEIA but detectable by ELISA. 4, 5 To address this discrepancy, we analysed sera from six patients with PV with undetectable anti-Dsg3 or anti-Dsg1 autoantibodies by CLEIA.The patients’ characteristics are shown in Figure 1 (a, b). This study was conducted in accordance with the Declaration of Helsinki principles and all participants gave written informed consent. The clinical information of case 2 was previously reported by us. 5 Sera from cases 3, 5 and 6 were previously used for the study of Fujio et al. 4 All cases were diagnosed as PV. Serum from case 1 was collected when case 1 had relapse of oral erosions, and serum from case 5 was collected when they were in a remission phase. The other cases were in an active phase of PV. All sera failed to react with Dsg3 or Dsg1 in CLEIA (STACIA MEBLux test Dsg3 and Dsg1; MBL, Tokyo, Japan)(Fig. 1 a, b). However, anti-Dsg1 or anti-Dsg3 autoantibodies were detected by ELISA (Mesacup-2 Desmoglein Test Dsg3 and Dsg1; MBL)(Fig. 1a, b), where the same Dsg3 and Dsg1 recombinant proteins were used in both CLEIA and ELISA. All cases had IgG autoantibodies reacting to intercellular spaces of the normal human epidermis (data not shown). These results confirmed that these sera contained anti-Dsg3 or anti-Dsg1 autoantibodies that were detectable by ELISA but not by CLEIA. Therefore, to address the discrepancy between the results of these assays, we focused on the great difference in incubation time for antigen–antibody reaction between CLEIA