Development and clinical application of a highly sensitive serological assay for detecting thrombospondin type-1 domain-containing 7A antibodies in idiopathic membranous nephropathy

IF 1.7 4区医学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of immunological methods Pub Date : 2026-05-01 Epub Date: 2026-05-19 DOI:10.1016/j.jim.2026.114065

Tianyu Zheng , Xuanli Tang , Shangbin Kao , Nannan Tian , Yufeng Yan , Dian Li , Canlong Yan , Yuan Qin , Xiumei Zhou , Yigang Wang , Guoqin Wang , Biao Huang

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引用次数: 0

Abstract

Objective

To establish a highly sensitive serological assay for detecting serum thrombospondin type-1 domain-containing 7 A antibodies (THSD7A-Ab) in patients with idiopathic membranous nephropathy (IMN), thereby addressing the current gap in available diagnostic biomarkers.

Methods

A time-resolved fluorescence immunoassay (TRFIA) was established by immobilizing THSD7A protein on the solid phase, which captured THSD7A-Ab present in serum samples. Detection was performed using a europium (Eu³⁺)-labeled secondary antibody. Serum THSD7A-Ab levels were measured in both THSD7A-positive IMN patients and M-type phospholipase A2 receptor (PLA2R)-negative IMN patients, and the corresponding seropositivity rates were determined.

Results

The TRFIA-THSD7A-Ab assay demonstrated a sensitivity of 0.13 RU/mL, with a linear range from 0.13 to 600 RU/mL. The intra-assay precision ranged from 3.35% to 4.63%, while the inter-assay precision ranged from 5.21% to 9.07%. The recovery rate was 93.78% to 107.20%. Serum THSD7A-Ab levels in THSD7A-associated IMN patients (176.54 ± 224.77 RU/mL) were significantly higher PLA2R-associated IMN patients (11.38 ± 4.76 RU/mL, P < 0.0001) and healthy controls (7.86 ± 2.92 RU/Ml, P < 0.0001). The cut-off value for distinguishing THSD7A-associated IMN was determined to be 13.70 RU/mL. Among PLA2R-negative IMN patients, the positive rate of THSD7A-Ab was 1.6%.

Conclusion

A highly sensitive TRFIA method for detecting serum THSD7A-Ab was successfully established for the first time, demonstrating excellent discriminative capability for identifying THSD7 A-positive patients.

查看原文本刊更多论文

一种检测特发性膜性肾病中含血小板反应蛋白1型结构域7A抗体的高灵敏度血清学方法的开发和临床应用。

目的：建立一种高灵敏度的血清学检测方法，用于检测特发性膜性肾病（IMN）患者血清中含有7 a抗体（THSD7A-Ab）的血小板反应蛋白1型结构域，从而解决目前可用诊断生物标志物的空白。方法：将THSD7A蛋白固定在固相上，建立时间分辨荧光免疫法（TRFIA），捕获血清样品中的THSD7A- ab。采用铕（Eu3+）标记的二抗进行检测。测定thsd7a阳性IMN患者和m型磷脂酶A2受体（PLA2R）阴性IMN患者血清中THSD7A-Ab水平，并测定相应的血清阳性率。结果：TRFIA-THSD7A-Ab检测灵敏度为0.13 RU/mL，线性范围为0.13 ~ 600 RU/mL。法内精密度为3.35% ~ 4.63%，法间精密度为5.21% ~ 9.07%。回收率为93.78% ~ 107.20%。thsd7a相关IMN患者血清THSD7A-Ab水平（176.54 ± 224.77 RU/mL）显著高于pla2r相关IMN患者（11.38 ± 4.76 RU/mL, P ）结论：首次成功建立了一种高灵敏度的TRFIA检测血清THSD7A-Ab的方法，对THSD7 A阳性患者具有良好的鉴别能力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of immunological methods 医学-免疫学

CiteScore

4.10

自引率

0.00%

发文量

120

审稿时长

3 months

期刊介绍： The Journal of Immunological Methods is devoted to covering techniques for: (1) Quantitating and detecting antibodies and/or antigens. (2) Purifying immunoglobulins, lymphokines and other molecules of the immune system. (3) Isolating antigens and other substances important in immunological processes. (4) Labelling antigens and antibodies. (5) Localizing antigens and/or antibodies in tissues and cells. (6) Detecting, and fractionating immunocompetent cells. (7) Assaying for cellular immunity. (8) Documenting cell-cell interactions. (9) Initiating immunity and unresponsiveness. (10) Transplanting tissues. (11) Studying items closely related to immunity such as complement, reticuloendothelial system and others. (12) Molecular techniques for studying immune cells and their receptors. (13) Imaging of the immune system. (14) Methods for production or their fragments in eukaryotic and prokaryotic cells. In addition the journal will publish articles on novel methods for analysing the organization, structure and expression of genes for immunologically important molecules such as immunoglobulins, T cell receptors and accessory molecules involved in antigen recognition, processing and presentation. Submitted full length manuscripts should describe new methods of broad applicability to immunology and not simply the application of an established method to a particular substance - although papers describing such applications may be considered for publication as a short Technical Note. Review articles will also be published by the Journal of Immunological Methods. In general these manuscripts are by solicitation however anyone interested in submitting a review can contact the Reviews Editor and provide an outline of the proposed review.