Specific chemoluminescence and immunoasdorption tests for anti-DFS70 antibodies avoid false positive results by indirect immunofluorescence

Bizzaro, Nicola; Tonutti, Elio; Tampoia, Marilina; Infantino, María; Cucchiaro, Francesco; Pesente, Fiorenza; Morozzi, Gabriella; Fabris, Martina; Villalta, Danilo

doi:10.1016/j.cca.2015.10.008

Cited by 43 publications

(42 citation statements)

References 32 publications

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“…It was therefore necessary to distinguish these NFS sera from the DFS sera by conducting confirmatory tests to validate the presence of autoantibodies to DFS70/LEDGFp75 in the DFS sera and their absence in the NFS sera. We found a strong concordance in the detection of anti-DFS70/LEDGFp75 by the different confirmatory tests in the DFS sera, highlighting the growing consensus that confirmation of a positive DFS pattern by the ANA IIF-HEp-2 test should be adequately achieved with additional tests [4, 5, 7, 21, 22, 24, 25]. However, we suggest that confirmatory WB data should be interpreted with caution given that whole lysates prepared from different cell lines may have varying expression levels of the DFS70/LEDGFp75 protein [19, 20], which could lead to inter-laboratory variations in DFS serum reactivity if the same cell line is not universally used.…”

Section: Discussionsupporting

confidence: 54%

“…To ensure the accurate detection of these autoantibodies, we initially tested for their presence using the HEp-2 based ANA-IIF test and then confirmed the results using three different but complementary detection platforms. These confirmatory tests were necessary in light of growing concerns that the recognition of the DFS pattern by the ANA-IIF test still remains challenging in the clinical laboratory setting [7, 21–25]. …”

Section: Discussionmentioning

confidence: 99%

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Detection of autoantibodies to DSF70/LEDGFp75 in Mexican Hispanics using multiple complementary assay platforms

Mercado

Gómez-Bañuelos

Navarro-Hernández

et al. 2016

Autoimmun Highlights

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PurposeAntinuclear autoantibodies (ANA) targeting the dense fine speckled antigen DFS70, also known as lens epithelium-derived growth factor p75 (LEDGF/p75), are attracting attention due to their low frequency in systemic rheumatic diseases but increased frequency in clinical laboratory referrals and healthy individuals (HI). These ANA specifically recognize the stress protein DFS70/LEDGFp75, implicated in cancer, HIV-AIDS, and inflammation. While their frequency has been investigated in various ethnic populations, there is little information on their frequency among Hispanics/Latinos. In this study, we determined the frequency of anti-DFS70/LEDGFp75 autoantibodies in Mexican Hispanics using multiple detection platforms.MethodsThe frequency of anti-DFS70/LEDGFp75 antibodies was determined in 171 individuals, including 71 dermatomyositis (DM) patients, 47 rheumatoid arthritis (RA) patients, 30 obesity (OB) patients, and 23 HI. Antibody detection was achieved using four complementary assay platforms: indirect immunofluorescence, Western blotting, ELISA, and chemiluminescent immunoassay.ResultsWe detected relatively low frequencies of anti-DFS70/LEDGFp75 antibodies in patients with DM (1.4%), RA (4.3%), and OB (6.6%), and elevated frequency (17.4%) in HI. A strong concordance between the different antibody detection platforms was observed.ConclusionsThe low frequency of anti-DFS70/LEDGFp75 antibodies in Mexican patients with rheumatic diseases, but relatively higher frequency in HI, is consistent with previous observations with non-Hispanic populations, suggesting that geographic differences or ethnicity do not influence the frequency of these autoantibodies. Our results also highlight the importance of confirmatory assays for the accurate detection of these autoantibodies. Future studies with larger cohorts of healthy Hispanics/Latinos are needed to confirm if their anti-DFS70/LEDGFp75 antibody frequencies are significantly higher than in non-Hispanics.

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Section: Discussionsupporting

confidence: 54%

Section: Discussionmentioning

confidence: 99%

Detection of autoantibodies to DSF70/LEDGFp75 in Mexican Hispanics using multiple complementary assay platforms

Mercado

Gómez-Bañuelos

Navarro-Hernández

et al. 2016

Autoimmun Highlights

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“…Based on this trend, the complexity and heterogeneity of ANA screening populations change significantly from one clinical lab to the other. Systematic review for DFS70 autoantibody positivity has been performed by multiple groups [4, 6, 30]. The majority of clinical studies have used IIF-HEp2 for establishing a suspicion of DFS70 pattern and the rates of positivity for DFS70 autoantibodies in each group varied widely between studies [7, 9, 18, 21, 28, 31–44].…”

Section: Prevalence Of Dfs70 Autoantibodiesmentioning

confidence: 99%

“…The majority of clinical studies have used IIF-HEp2 for establishing a suspicion of DFS70 pattern and the rates of positivity for DFS70 autoantibodies in each group varied widely between studies [7, 9, 18, 21, 28, 31–44]. DFS70 antibodies have been reported in high titers from cohorts of healthy individuals, blood donors, patients being screened for ANA, patients with various autoimmune disorders and various non-autoimmune disorders including cancers [4, 6, 30]. These studies have shown that DFS70 autoantibodies lack distinct clinical association, with most disease groups, except for certain inflammatory conditions of eyes and skin [4, 6, 7, 18, 30, 44, 45].…”

Section: Prevalence Of Dfs70 Autoantibodiesmentioning

confidence: 99%

“…DFS70 antibodies have been reported in high titers from cohorts of healthy individuals, blood donors, patients being screened for ANA, patients with various autoimmune disorders and various non-autoimmune disorders including cancers [4, 6, 30]. These studies have shown that DFS70 autoantibodies lack distinct clinical association, with most disease groups, except for certain inflammatory conditions of eyes and skin [4, 6, 7, 18, 30, 44, 45]. The method of screening, selection, and composition of study cohorts may also influence the reported rates of DFS70 autoantibody positivity.…”

Section: Prevalence Of Dfs70 Autoantibodiesmentioning

confidence: 99%

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Analysis of DFS70 pattern and impact on ANA screening using a novel HEp-2 ELITE/DFS70 knockout substrate

Malyavantham

2017

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Indirect immunofluorescence (IIF) using human epithelial cell (HEp-2) substrate is a widely used and the recommended method for screening of antinuclear antibodies (ANA). Dense fine speckled (DFS70) pattern on HEp-2 has been widely reported in various healthy and disease groups. Interpretation of DFS70 pattern can be challenging on a conventional HEp-2 substrate due to its similarity to some of the disease associated patterns. The high prevalence of DFS70 autoantibodies in normal population, lack of association with a particular disease group and a general negative association with systemic and ANA associated autoimmune rheumatic diseases (SARD/AARD) necessitates the confirmation of DFS70 pattern. Results using available commercial assays for confirmation of DFS70 autoantibodies do not always agree with IIF screening results further complicating the lab work flow and ANA algorithms. In this review, we discuss the prevalence of DFS70 antibodies and factors affecting the performance of IIF and DFS70 specific confirmatory assays. Factors that contribute to disagreement between DFS70 suspicion by IIF and confirmatory assays will also be discussed. In addition, we also describe a novel IIF HEp-2 substrate, and its positive impact on DFS70 reporting and ANA screening-confirmation algorithm.

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Dense fine speckled nuclear immunofluorescence: A mildly reassuring antinuclear antibody pattern meriting consideration

Fijałkowska,

Schwartz,

Woźniacka

2023

Immunity Inflam & Disease

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IntroductionAntinuclear antibodies (ANAs) are regarded as a hallmark of connective tissue diseases (CTDs) and play a key role in their diagnosis, but the value of some particular antibodies in management of patients and the disease prognosis is controversial. The mechanism underlying the production of ANAs in CTDs, other chronic inflammatory conditions and even in healthy people, is not completely elucidated. Anti‐DFS70 antibodies connected with the dense fine speckled autoantigen of 70 kD, known as the lens epithelium‐derived growth factor p75, are a subgroup of ANAs. Their presence and coexistence with other antibodies and their clinical significance are the matter of debate.MethodsBased on literature data, the authors focused on current knowledge explaining the role of anti‐DFS70 antibodies in selected CTDs.ResultsHowever, the literature data is ambiguous and does not fully support the validity of the anti‐DFS70 assay for a specific CTD diagnosis. Most researchers claim that the presence of anti‐DFS70 as the only one usually exclude the diagnosis of CTD. Nevertheless, its coexistence with other ANAs is not an excluding factor but has predictive value due to more favorable course of CTD. Such situations may also suggest an enhanced risk of the development of a CTD in the future.ConclusionsAlthough more studies are needed in this field, it seems reasonable to ascertain the presence of anti‐DFS70 in routine clinical practice.

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Specific chemoluminescence and immunoasdorption tests for anti-DFS70 antibodies avoid false positive results by indirect immunofluorescence

Cited by 43 publications

References 32 publications

Detection of autoantibodies to DSF70/LEDGFp75 in Mexican Hispanics using multiple complementary assay platforms

Detection of autoantibodies to DSF70/LEDGFp75 in Mexican Hispanics using multiple complementary assay platforms

Analysis of DFS70 pattern and impact on ANA screening using a novel HEp-2 ELITE/DFS70 knockout substrate

Dense fine speckled nuclear immunofluorescence: A mildly reassuring antinuclear antibody pattern meriting consideration

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