Diagnostic profile on the IFA 40: HEp-20-10 – an immunofluorescence test for reliable antinuclear antibody screening

Rohwäder, Edda; Locke, Michael; Fraune, J.; Fechner, Kaï

doi:10.1586/14737159.2015.993612

Cited by 6 publications

(5 citation statements)

References 50 publications

(54 reference statements)

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“…Of note, the analytical performance of HEp-2 IIF crucially depends on the quality of the substrate and assay, particularly with regard to the ratio of mitotic versus interphase cells, cell morphology, reproducibility, and dilution linearity. The HEp-20-10 substrate (a variant of the standard HEp-2 cell line with a 10-fold higher number of mitotic cells), as used in the present study, has been reported to facilitate the recognition, discrimination and titer estimation of ANA patterns ( 63 ).…”

Section: Discussionmentioning

confidence: 99%

Recognition and Relevance of Anti-DFS70 Autoantibodies in Routine Antinuclear Autoantibodies Testing at a Community Hospital

Carter

Saschenbrecker

et al. 2018

Front. Med.

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Antinuclear autoantibodies (ANA) displaying a dense fine speckled pattern (DFS, ICAP AC-2) on HEp-2 cells are frequently observed in clinical laboratory referrals, often associated with anti-DFS70 specificity. Anti-DFS70 positive patients rarely develop systemic autoimmune rheumatic disease (SARD), especially in the absence of clinical evidence or additional anti-extractable nuclear antigen (ENA) antibodies, prompting suggestions that an isolated DFS70-specific ENA may be an exclusionary finding for SARD. In this study, the frequency and diagnostic significance of anti-DFS70 autoantibodies was investigated in a community hospital cohort of patients undergoing routine ANA testing. ANA screening was performed by HEp-20-10-based indirect immunofluorescence, followed by ENA profiling using a multiparametric line immunoassay (LIA). Of 6,511 patient samples tested for ANA in 2016, the DFS pattern was identified in 1,758 (27.0%), 720 (41.0%) of which were anti-DFS70 positive by LIA. Of these, 526 (73.1%) revealed isolated anti-DFS70 reactivity, while 194 (26.9%) showed additional ENA specificities. Among 1,038 anti-DFS70 negative or borderline samples, 778 (75.0%) were ENA profile negative, while the remaining 260 (25.0%) showed a varied presence of other ENA specificities. Chart reviews of patients with an isolated anti-DFS70 ANA affirmed that ANA-related SARD is rare in the absence of clinical evidence or other ENA specificities, there being no case thus far identified. Rheumatoid arthritis patients occasionally had an isolated anti-DFS70 ANA and were positive for rheumatoid factor and anti-cyclic citrullinated peptide antibodies. In conclusion, the recognition of a DFS ANA pattern using a mitotic-rich HEp-2 substrate, followed by confirmation of anti-DFS70 specificity should be a routine ANA testing service. Use of an expanded ENA profile and clinical correlation is necessary to affirm the “isolation” of anti-DFS70 as the cause of an ANA. Recognition of isolated anti-DFS70 ANA enables reassurance of patients that SARD is unlikely, thus avoiding referral for more extensive testing. The presence of significant elevations of other ENAs may reflect SARD and warrants close clinical correlation and follow-up.

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Section: Discussionmentioning

confidence: 99%

Recognition and Relevance of Anti-DFS70 Autoantibodies in Routine Antinuclear Autoantibodies Testing at a Community Hospital

Carter

Saschenbrecker

et al. 2018

Front. Med.

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“…Constant quality of the substrate is ensured by quality control measures throughout the production of the HEp-20-10-coated biochips. Reliability of the assay has been demonstrated by validation studies ( 57 ).…”

Section: Discussionmentioning

confidence: 99%

“…Samples and slides were processed using the Sprinter XL (Euroimmun AG, Lübeck, Germany), followed by automated evaluation using the EUROPattern Suite (Euroimmun AG, Lübeck, Germany). ANA detection was performed by the IFA 40: HEp-20-10 kit assay according to the manufacturer’s instructions (Euroimmun AG, Lübeck, Germany) ( 57 ). The IFA 40: HEp-20-10 allows for easier evaluation due to an increased spectrum of cells in the mitotic phase and of human nuclear antigens when compared to traditional Hep-2 cells.…”

Section: Methodsmentioning

confidence: 99%

Automated Processing and Evaluation of Anti-Nuclear Antibody Indirect Immunofluorescence Testing

et al. 2018

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Indirect immunofluorescence (IIF) is considered by the American College of Rheumatology (ACR) and the international consensus on ANA patterns (ICAP) the gold standard for the screening of anti-nuclear antibodies (ANA). As conventional IIF is labor intensive, time-consuming, subjective, and poorly standardized, there have been ongoing efforts to improve the standardization of reagents and to develop automated platforms for assay incubation, microscopy, and evaluation. In this study, the workflow and performance characteristics of a fully automated ANA IIF system (Sprinter XL, EUROPattern Suite, IFA 40: HEp-20-10 cells) were compared to a manual approach using visual microscopy with a filter device for single-well titration and to technologist reading. The Sprinter/EUROPattern system enabled the processing of large daily workload cohorts in less than 8 h and the reduction of labor hands-on time by more than 4 h. Regarding the discrimination of positive from negative samples, the overall agreement of the EUROPattern software with technologist reading was higher (95.6%) than when compared to the current method (89.4%). Moreover, the software was consistent with technologist reading in 80.6–97.5% of patterns and 71.0–93.8% of titers. In conclusion, the Sprinter/EUROPattern system provides substantial labor savings and good concordance with technologist ANA IIF microscopy, thus increasing standardization, laboratory efficiency, and removing subjectivity.

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“…Monkey liver tissue is helpful to determine fluorescence patterns, especially in some undistinguishable patterns. The fluorescence characteristics in monkey liver tissue are key points of identification, thus avoiding mistakes ( 23 , 24 ). In the first incubation step, specific antibodies from the diluted patient sample bind to the solid-phase bound antigens.…”

Section: Methodsmentioning

confidence: 99%

The presence of autoantibodies is associated with improved overall survival in lung cancer patients

Jing,

Zhao,

Cai

et al. 2023

Front. Oncol.

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ObjectiveAutoantibodies have been reported to be associated with cancers. As a biomarker, autoantibodies have been widely used in the early screening of lung cancer. However, the correlation between autoantibodies and the prognosis of lung cancer patients is poorly understood, especially in the Asian population. This retrospective study investigated the association between the presence of autoantibodies and outcomes in patients with lung cancer.MethodsA total of 264 patients diagnosed with lung cancer were tested for autoantibodies in Henan Provincial People’s Hospital from January 2017 to June 2022. The general clinical data of these patients were collected, and after screening out those who met the exclusion criteria, 151 patients were finally included in the study. The Cox proportional hazards model was used to analyze the effect of autoantibodies on the outcomes of patients with lung cancer. The Kaplan-Meier curve was used to analyze the relationship between autoantibodies and the overall survival of patients with lung cancer.ResultsCompared to lung cancer patients without autoantibodies, those with autoantibodies had an associated reduced risk of death (HRs: 0.45, 95% CIs 0.27~0.77), independent of gender, age, smoking history, pathological type, and pathological stage of lung cancer. Additionally, the association was found to be more significant by subgroup analysis in male patients, younger patients, and patients with small cell lung cancer. Furthermore, lung cancer patients with autoantibodies had significantly longer survival time than those without autoantibodies.ConclusionThe presence of autoantibodies is an independent indicator of good prognosis in patients with lung cancer, providing a new biomarker for prognostic evaluation in patients with lung cancer.

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Diagnostic profile on the IFA 40: HEp-20-10 – an immunofluorescence test for reliable antinuclear antibody screening

Cited by 6 publications

References 50 publications

Recognition and Relevance of Anti-DFS70 Autoantibodies in Routine Antinuclear Autoantibodies Testing at a Community Hospital

Recognition and Relevance of Anti-DFS70 Autoantibodies in Routine Antinuclear Autoantibodies Testing at a Community Hospital

Automated Processing and Evaluation of Anti-Nuclear Antibody Indirect Immunofluorescence Testing

The presence of autoantibodies is associated with improved overall survival in lung cancer patients

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