检验医学 ›› 2023, Vol. 38 ›› Issue (12): 1135-1140.DOI: 10.3969/j.issn.1673-8640.2023.12.005

• 自身免疫性疾病生物标志物研究与应用 • 上一篇    下一篇

胞质型抗核抗体对间接免疫荧光法检测抗中性粒细胞胞质抗体的影响

杨晓, 李恩灵, 吴丽霞, 戴颖欣, 王之青, 黄昊, 郑冰()   

  1. 上海交通大学医学院附属仁济医院检验科,上海 200127
  • 收稿日期:2023-09-18 修回日期:2023-11-07 出版日期:2023-12-30 发布日期:2024-02-20
  • 通讯作者: 郑 冰,E-mail:zhengbing@renji.com
  • 作者简介:郑 冰,E-mail:zhengbing@renji.com
    杨 晓,女,1988年生,硕士,主管技师,主要从事自身抗体检测工作。
  • 基金资助:
    上海市扬帆计划资助项目(22YF1438400);广东省钟南山基金资助项目(ZNSXS-20220007)

Influence of cytoplasmic antinuclear antibody on indirect immunofluorescence assay based antineutrophil cytoplasmic antibody determination

YANG Xiao, LI Enling, WU Lixia, DAI Yingxin, WANG Zhiqing, HUANG Hao, ZHENG Bing()   

  1. Department of Clinical Laboratory,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200127,China
  • Received:2023-09-18 Revised:2023-11-07 Online:2023-12-30 Published:2024-02-20

摘要:

目的 探讨胞质型抗核抗体(ANA)对间接免疫荧光法(IFA)检测抗中性粒细胞胞质抗体(ANCA)的影响。方法 收集2023年6—7月上海交通大学医学院附属仁济医院非ANCA相关性血管炎,且ANA荧光模型为胞质型的患者血清样本66例,其中胞质线性/肌动蛋白型(AC-15型)2例、胞质丝状/微管型(AC-16型)4例、胞质散点型(AC-18型)7例、胞质致密颗粒型(AC-19型)25例、胞质细颗粒型(AC-20型)13例、胞质网状/线粒体样型(AC-21型)15例。采用IFA检测ANCA。采用免疫印迹法检测抗可溶性核抗原(ENA)抗体谱和自身免疫性肝病(AILD)抗体谱。采用酶联免疫吸附试验(ELISA)检测抗双链DNA(dsDNA)抗体、抗核小体抗体和ANCA谱。结果 66例胞质型ANA阳性样本中,有65例(98.5%)ANCA为阴性,1例(1.5%)为不典型胞质型抗中性粒细胞胞质抗体(cANCA),ANCA谱均为阴性。采用IFA检测66例胞质型ANA阳性样本的ANCA,乙醇固定基质片有42.4%(28/66)为ANCA阴性,有30.3%(20/66)为不典型ANCA,有22.7%(15/66)为核周型抗中性粒细胞胞质抗体(pANCA),有4.5%(3/66)为cANCA;甲醛固定基质片仅1例(1.5%)阳性。25例AC-19型阳性样本中,有24例(96%)乙醇固定基质片表现为阳性,其中12例(48.0%)为pANCA,11例(44.0%)为非典型ANCA,1例(4.0%)为不典型cANCA;合并抗核糖体P蛋白(Rib.P)抗体阳性时,pANCA的阳性率(64.7%)显著高于抗Rib.P抗体阴性时(12.5%)(P=0.02)。15例AC-21型阳性样本中,有6例(40.0%)乙醇固定基质片表现为阴性中,有6例(40.0%)为非典型ANCA,2例(13.3%)为cANCA,1例(6.7%)为pANCA。AC-15型、AC-16型、AC-18型和AC-20型阳性样本乙醇固定基质片表现均以阴性为主,表现为阳性的样本多合并抗ENA抗体谱阳性。结论 采用IFA检测ANCA时,胞质型ANA对甲醛固定基质片的影响较小,对乙醇固定基质片的干扰多见于AC-19型和AC-21型。当AC-19型和AC-21型合并抗ENA抗体阳性时,可产生pANCA或非典型ANCA干扰。

关键词: 抗核抗体, 胞质型, 抗中性粒细胞胞质抗体, 干扰

Abstract:

Objective To investigate the influence of cytoplasmic antinuclear antibody(ANA)on indirect immunofluorescence assay(IFA) based antineutrophil cytoplasmic antibody(ANCA)determination. Methods Totally,66 patients with non-ANCA-associated vasculitis with ANA patterns of cytoplasmic fibrillar linear(AC-15,2 cases),cytoplasmic fibrillar filamentous(AC-16,4 cases),cytoplasmic discrete dots/GW body-like(AC-18,7 cases),cytoplasmic dense fine speckled(AC-19,25 cases),cytoplasmic fine speckled(AC-20,13 cases)and cytoplasmic reticular/AMA(AC-21,15 cases)were enrolled from Renji Hospital of Shanghai Jiao Tong University School of Medicine from June to July 2023. IFA was carried out for ANCA,western blot was carried out for anti-extractable nuclear antigen(ENA)antibody spectrum and anti-autoimmune liver disease(AILD)antibody spectrum,and enzyme-linked immunosorbent assay(ELISA) was used for anti-double-stranded DNA(dsDNA) antibody,anti-nucleosome antibody and ANCA spectrum. Results Among 66 cases of cytoplasmic ANA,98.5%(65/66)were ANCA negative,and 1 case(1.5%) of atypical cytoplasmic ANA(cANCA)was ANCA negative. In ethanol-fixed human neutrophils,42.4%(28/66) were negative,30.3%(20/66)were atypical ANCA,22.7%(15/66)were perinuclear ANCA(pANCA),and 4.5%(3/66)were cANCA. The positive rate of formalin fixed neutrophils was 1.5%(1/66). The 96%(24/25)of AC-19 samples were positive in ethanol-fixed substrate,48.0%(12/25)for pANCA,44.0%(11/25)for atypical ANCA,and 4.0%(1/25) for atypical cANCA. The positive rate of pANCA was higher when accompanied with anti-ribosomal P protein antibody than anti-ribosomal protein(Rib. P) antibody negative(64.7%,12.5%,P=0.02). AC-21 mainly showed as negative and atypical ANCA in ethanol-fixed substrate,whose percentages were both 40.0%(6/15),13.3%(2/15) for cANCA,and 6.7%(1/15) for pANCA. AC-15,AC-16,AC-18 and AC-20 were mainly negative in ethanol-fixed substrate,and most of the positive samples in ethanol-fixed substrate were positive when accompanied with anti-ENA antibody spectrum. Conclusions Cytoplasmic ANA has little interference on formalin fixed neutrophils in IFA based ANCA determination,and the interference of cytoplasmic ANA in ethanol-fixed neutrophils mostly shows in AC-19 and AC-21. Interferences showing as pANCA or atypical ANCA may occur when these 2 ANA patterns combining with other nuclear antigens.

Key words: Antinuclear antibody, Cytoplasmic type, Antineutrophil cytoplasmic antibody, Interference

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