Verizhnikova Zh G, Aleksandrova E N, Novikov A A, Panafidina T A, Seredavkina N V, Popkova T V, Aizina N L, Nasonov E L
Klin Lab Diagn. 2017 Mar;62(3):173-7.
Thew antinuclear antibodies (ANA) consist heterogeneous group of auto antibodies reacting with various components of nucleus and cytoplasm. The ANA is a main serological marker of systemic lupus erythematosus (SLE). The implementation in clinical practice of new highly productive techniques of immune analysis using automated systems sets up prerequisites for standardization and amelioration of reproducibility of detection of ANA. The study was carried out to compare diagnostic significance of automated techniques of screening detection of ANA (indirect immunofluorescence test on cells HEp-2 (IIFT-HEp-2)), enzyme-linked immunosorbent assay (ELISA) and multi-complex immune analysis (MIA, using suspension technology xMAP) in serum of patients with SLE. The serums from 94 patients with SLE were analyzed. The comparison group included 70 patients with other rheumatic diseases. The control group consisted of 30 healthy donors. The screening detection of ANA using technique IIFT-HEp-2 was implemented on automated platform AKLIDES, ELISA - on automated analyzer ALEGRIA and MIA on automated analyzer BioPlex 2200. The technique IIFT-HEp-2 demonstrated the most high diagnostic sensitivity as compared with ELISA and MIA- BioPlex 2200 (96.8%; 79.8% and 82.9% correspondingly). The general diagnostic specificity of detection of ANA using technique IIFT-HEp-2 was lower than in case of ELISA and MIO-BioPlex 2200 (40%, 70% and 57% correspondingly). In the group of healthy donors the lowest diagnostic specificity was observed in ANA screening analysis using MIA-BioPlex 2200 (80%) while in case of applying IIFT-HEp-2 and ELISA indices of diagnostic specificity made up 93.3% and 96.7% correspondingly. The ANA analysis of mix of 26 nuclear antigens using ELISA technique was a reliable laboratory test for diagnostic of SLE (likelihood ratio of positive result - 2.66). By the level of likelihood ratio of negative result of the IIFT-HEp-2 technique was more informative test for exclusion of diagnosis of SLE than techniques of ELISA and MIA-BioPlex 2200 (0.08; 0.29 and 0.3 correspondingly). The detection of ANA using technique of is the most preferable primary screening test for diagnostic of SLE. The ELISA of antibodies to mix of nuclear antigens and MIA on the basis of xMAP technology are less preferable screening tests for diagnostic of SLE as compared with IIFT-HEp-2 because of false-negative results in 20% and 17% of cases correspondingly. ELISA and MIA are to applied as confirmatory screening tests permitting to detect antigen-specific ANA in patients with SLE with positive results of IIFT-HEp-2.
抗核抗体（ANA）由与细胞核和细胞质的各种成分发生反应的异质性自身抗体组成。ANA是系统性红斑狼疮（SLE）的主要血清学标志物。使用自动化系统的新型高效免疫分析技术在临床实践中的应用为ANA检测的标准化和提高重现性奠定了前提条件。本研究旨在比较ANA筛查检测的自动化技术（HEp-2细胞间接免疫荧光试验（IIFT-HEp-2））、酶联免疫吸附测定（ELISA）和多重复合免疫分析（MIA，使用悬浮技术xMAP）在SLE患者血清中的诊断意义。分析了94例SLE患者的血清。对照组包括70例其他风湿性疾病患者。对照组由30名健康供者组成。使用IIFT-HEp-2技术在自动化平台AKLIDES上进行ANA的筛查检测，使用ELISA在自动化分析仪ALEGRIA上进行检测，使用MIA在自动化分析仪BioPlex 2200上进行检测。与ELISA和MIA-BioPlex 2200相比，IIFT-HEp-2技术显示出最高的诊断敏感性（分别为96.8%、79.8%和82.9%）。使用IIFT-HEp-2技术检测ANA的总体诊断特异性低于ELISA和MIO-BioPlex 2200（分别为40%、70%和57%）。在健康供者组中，使用MIA-BioPlex 2200进行ANA筛查分析时观察到最低的诊断特异性（80%），而使用IIFT-HEp-2和ELISA时诊断特异性指标分别为93.3%和96.7%。使用ELISA技术对26种核抗原混合物进行ANA分析是诊断SLE的可靠实验室检测方法（阳性结果的似然比为2.66）。就阴性结果的似然比而言，IIFT-HEp-2技术比ELISA和MIA-BioPlex 2200技术更有助于排除SLE诊断（分别为0.08、0.29和0.3）。使用IIFT-HEp-2技术检测ANA是诊断SLE最优选的初筛试验。与IIFT-HEp-2相比，基于xMAP技术的核抗原混合物抗体ELISA和MIA作为SLE诊断的筛查试验不太优选，因为分别有20%和17%的病例出现假阴性结果。ELISA和MIA应用于确证性筛查试验，用于检测IIFT-HEp-2结果为阳性的SLE患者中的抗原特异性ANA。
