抗核抗体筛查试验的新方法：一种检测结缔组织病特征性抗核抗体的酶联免疫吸附测定法

[Newer approach of screening test for antinuclear antibodies: an enzyme-linked immunosorbent assay detecting antinuclear antibodies characteristic of connective tissue diseases].

作者信息

Asanuma H, Miyake J, Miyawaki S

机构信息

Clinical Laboratory and Rheumatic Disease Center, Center for Adult Disease, Kurashiki.

出版信息

Nihon Rinsho Meneki Gakkai Kaishi. 1997 Oct;20(5):417-27. doi: 10.2177/jsci.20.417.

DOI:10.2177/jsci.20.417

PMID:9391305

Abstract

An enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of antinuclear antibodies (ANAs) previously established as diagnostic and/or prognostic marker ANAs for various connective tissue diseases. The antigen used in ELISA is a mixture of purified recombinant or natural antigens including single-and double-stranded DNA, RNP, Sm, SS-A/Ro, SS-B/La, centromere, topoisomerase I and Jo-1 antigens. Thirty hundred and fifty nine patients sera from a variety of connective tissue diseases and 113 normal human sera (NHS) were examined. ELISA ANAs were positive in 3.5% of NHS and 80.2% of patients sera at cut off index 11.5, whereas indirect immunofluorescent antinuclear antibodies (FANAs) using HEp-2 cells were positive in 9.7% of NHS and 92.5% of patients sera at 1:160 serum dilution. More than 80% of sera from systemic lupus erythematosus, mixed connective tissue disease and primary Sjögrens disease were ELISA ANAs positive. Mean value of ELISA ANAs was highest in sera of patients with MCTD. ELISA ANAs were positive in 92.5% of sera with marker ANAs for connective tissue diseases. Mean value of ELISA ANAs was higher in sera with more than two marker ANAs than in sera with a single ANA or in sera without marker ANAs. In contrast incidence and mean value of ELISA ANAs were low in sera positive for anti topoisomerase I antibody or anti Jo-1 antibody. Sensitivity, specificity and agreement (accuracy) for connective tissue diseases with marker ANAs were as follows: ELISA ANAs (at index 11.5): 92.5%, 88.3% and 90.9%: FANAs (at 1:160 serum dilution): 99.0%, 70.4% and 88.1%, respectively. ELISA ANAs, thus, are specific for connective tissue diseases when compared to FANAs and previous ELISA for the detection of total ANAs. Moreover, ELISA ANAs are able to measure precise ANAs titers and are much less labor intensive when screening a large number of clinical specimens.

摘要

已开发出一种酶联免疫吸附测定（ELISA）法，用于检测抗核抗体（ANA），此前已确定其为多种结缔组织疾病的诊断和/或预后标志物。ELISA中使用的抗原是纯化的重组或天然抗原的混合物，包括单链和双链DNA、核糖核蛋白（RNP）、史密斯（Sm）抗原、干燥综合征-A/罗（SS-A/Ro）抗原、干燥综合征-B/拉（SS-B/La）抗原、着丝粒、拓扑异构酶I和Jo-1抗原。检测了来自各种结缔组织疾病的359例患者血清和113份正常人血清（NHS）。在临界指数为11.5时，ELISA法检测ANA，NHS中有3.5%呈阳性，患者血清中有80.2%呈阳性；而使用人喉表皮样癌细胞（HEp-2细胞）的间接免疫荧光抗核抗体（FANA）在血清稀释度为1:160时，NHS中有9.7%呈阳性，患者血清中有92.5%呈阳性。系统性红斑狼疮、混合性结缔组织病和原发性干燥综合征患者血清中超过80%的ELISA法检测ANA呈阳性。混合性结缔组织病（MCTD）患者血清中ELISA法检测ANA的平均值最高。结缔组织病标志物ANA的血清中，92.5%的ELISA法检测ANA呈阳性。有两种以上标志物ANA的血清中ELISA法检测ANA的平均值高于只有一种ANA或无标志物ANA的血清。相比之下，抗拓扑异构酶I抗体或抗Jo-1抗体阳性的血清中ELISA法检测ANA的发生率和平均值较低。结缔组织病标志物ANA的敏感性、特异性和一致性（准确性）如下：ELISA法检测ANA（临界指数为11.5时）：分别为92.5%、88.3%和90.9%；FANA（血清稀释度为1:160时）：分别为99.0%、70.4%和88.1%。因此，与FANA及之前检测总ANA的ELISA法相比，ELISA法检测ANA对结缔组织病具有特异性。此外，ELISA法检测ANA能够精确测定ANA滴度，在筛查大量临床标本时劳动强度小得多。