ANA-specific antibodies, ANA patterns, anti-ds-DNA results, and clinical diagnosis: a laboratory and clinical audit

Anis, Sabiha; Fatima, Areej; Jabbar, Sidra Abdul; Arain, Tayyab

doi:10.1007/s12026-022-09347-z

Cited by 3 publications

(4 citation statements)

References 25 publications

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“…The deposition of various autoantibodies plays a key role in the onset, progression, and severity of LN and is of great value in its diagnosis, treatment, and prognosis ( Oelzner et al, 2003 ; van der Vlag and Berden, 2011 ). Anti-ds-DNA and ANA-specific antibodies are specific to systemic autoimmune diseases ( Anis et al, 2023 ), and ds-DNA-Ab was significantly positively correlated with c_Bacteria_unclassified in our study. SLEDAI is a commonly used disease activity scoring tool in clinical practice.…”

Section: Discussionsupporting

confidence: 59%

Synbiotic regulates gut microbiota in patients with lupus nephritis: an analysis using metagenomic and metabolome sequencing

Zhu,

Cui,

Liu

et al. 2024

Front. Microbiol.

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ObjectiveTo investigate the changes in gut microbes and their metabolites after administering synbiotics to patients with new-onset lupus nephritis (LN) treated using a conventional method and provide a theoretical basis for finding new targets for the diagnosis and treatment of LN.MethodsIn this study, a total of 12 participants were divided into the lupus and synbiotic groups. Stool samples and clinical data were collected before and after treatment for metagenomic, nontargeted metabolomic, and statistical analyses.ResultsThe relative abundances of the pathogenic bacteria Prevotella, Bacteroides, and Enterobacteriaceae_unclassified decreased after synbiotic treatment, whereas the abundances of Actinobacteria and Firmicutes increased. Further, the Firmicutes to Bacteroidetes ratio increased; however, the difference was not statistically significant (p > 0.05). α diversity analysis showed no significant differences in the intestinal microbial richness and diversity index of patients with LN between the groups before and after treatment (p > 0.05). β analysis showed the differences in the community structure between the samples of the two groups before and after treatment. Linear discriminant analysis effect size and receiver operating characteristic curve analyses revealed that Negativicutes (AUC = 0.9722) and Enterobacteriaceae_unclassified (AUC = 0.9722) were the best predictors of the lupus and synbiotic groups, respectively, before and after treatment. Joint analyses revealed that amino acid biosynthesis, aminoacyl-tRNA biosynthesis, purine metabolism, and other metabolic pathways may be involved in the changes in the metabolic function of patients with LN after the addition of synbiotics. Spearman’s correlation analysis revealed the interaction between clinical features and flora, and flora exhibited a complex biological network regulatory relationship.ConclusionSynbiotics regulate the metabolic functions of intestinal microorganisms in patients with LN and play a role in various biological functions. Synbiotic supplements may be safe and promising candidates for patients with LN.

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Section: Discussionsupporting

confidence: 59%

Synbiotic regulates gut microbiota in patients with lupus nephritis: an analysis using metagenomic and metabolome sequencing

Zhu,

Cui,

Liu

et al. 2024

Front. Microbiol.

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“…A retrospective analysis showed that, among 206 patients reactive for specific antibodies, 195 patients (95%) tested positive for ANAs. Within these cases, speckled and homogeneous were identified as the predominant ANA patterns [28]. A cross-sectional study among 495 SLE patients revealed that the most frequently observed ANA patterns were speckled (52.1%) and homogeneous (35.2%) patterns [29].…”

Section: Discussionmentioning

confidence: 99%

Discriminating between Homogeneous (AC-1) and Dense Fine Speckled (AC-2) Antinuclear Antibody Patterns: Re-Evaluation of Immunofluorescence Imaging

Yu,

Hsieh,

Huang

et al. 2023

Biomedicines

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Antinuclear antibodies (ANAs) are essential diagnostic markers in systemic autoimmune rheumatic diseases. Among the 30 ANA patterns, homogeneous (AC-1) and dense fine speckled (AC-2) should be focused on owing to their somewhat indistinct presentation in immunofluorescence imaging and distinct correlation with clinical conditions. This study aimed to develop a flowchart to guide discrimination between AC-1 and AC-2 patterns and to re-evaluate ANA samples according to this flowchart to verify its detection ability. We re-evaluated immunofluorescence imaging of 62 ANA blood samples simultaneously subjected to solid-phase assays for autoantibodies against dsDNA, nucleosomes, histones, and DFS70. The results showed statistically significant odd ratios (ORs) of detection of anti-DFS70 using AC-2 after re-evaluation of total samples (OR 101.9, 95% CI 11.7–886.4, p-value < 0.001) and subgroup analysis of patients’ samples (OR 53.8, 95% CI 5.9–493.6, p-value < 0.001). The OR of anti-nucleosome/histone/dsDNA detection using AC-1 in re-evaluated data increased to 5.43 (95% CI 1.00–29.61, p-value = 0.05). In the analysis of specific autoantibodies, more than half of the samples with an AC-2 pattern (54.2%) had specific autoantibodies other than anti-DFS70. We conclude that the flowchart for discriminating between AC-1 and AC-2 ANA patterns in this study is a viable practical guide for other laboratories when encountering equivocal ANA results.

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“…Similarly, Menor Almagro et al reported 69.6% association of speckled pattern with SS-A/Ro 60, SS-A/Ro52, and anti-La [30], followed by homogenous pattern (N=167) which was associated with dsDNA135 (80.8%), nucleosome 110 (65.9%), and histones 42 (25.1%). Again, Anis et al reported that speckled and homogenous were the predominant ANA patterns (56% and 42%, respectively) and they were associated with Sm/RNP, histones, and nucleosomes antibodies [12].…”

Section: Discussionmentioning

confidence: 99%

“…Several studies reported positive autoantibodies by different methods in ANA-IIF assay-negative SLE. In a study by Enis et al, 5% of his patients was negative for ANA-IIF assay and positive for at least one antibody by immunoblot assay [12]. Another study by Gualtierotti reported that 7.4% of their SLE patients were ANA-IIF negative at the beginning of follow-up [13].…”

Section: Introductionmentioning

confidence: 93%

Negative ANA-IIF in SLE patients: what is beyond?

et al. 2023

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The antinuclear antibody (ANA) test has high sensitivity in diagnosing and classifying systemic lupus erythematosus (SLE). Objectives To describe the immunological pattern of SLE patients through investigating specific antinuclear autoantibodies by enzyme dot immunoassay and studying their frequency in both positive and negative ANA indirect immunofluorescence assay (IIF) cases. Methods In a cross-sectional study, blood samples from 393 newly diagnosed SLE patients were analyzed using (IIF) on HEp-2 cells and ANA dot immunoassay by automated enzyme immunoassay (EIA) to detect 19 antibodies. Results Ninety-one percent of the patients are females; their mean age was 37 ± 12.28. Antinuclear antibody (ANA) was detected by IIF in 82.4% of cases, with 181 (46.1%) speckled and 167 (42.4%) homogeneous ANA patterns. The majority of patients (96%) demonstrated autoantibodies via EIA. Among the ANA-IIF-negative patients, 97.2% demonstrated autoantibodies. There was a significant difference in the frequency of certain autoantibodies between SLE patients with negative and positive ANA-IIF (1.44 0.73, 3.12 2.09, p = 0.00) respectively. Conclusion The results of analyzing 19 autoantibodies with the ANA staining pattern increased the significance of analyzing the immune profile even if IIF is negative when clinical symptoms strongly suggest SLE diagnosis. Certain autoantibodies may evade staining by the IFA approach while they are present in the patient’s serum, and they may not be detected by the ANA EIA profile if it does not contain that antigenic substrate. Key Points• Indirect immunofluorescence on Hep-2 is the conventional method for ANA detection and is regarded as the “gold standard” for testing in clinical practice for SLE.• In our study, ANA profile dot enzyme immunoassay (EIA)-based test was performed to evaluate 19 autoantibodies in SLE patients either positive or negative for ANA-IIF.• The presence of anti-dsDNA with ANA-IIF-negative serum in 32.4% of SLE patients provides evidence that not all anti-dsDNA antibodies are identified on standard HEp-2 substrates.• certain autoantibodies can evade staining by the ANA-IIF method despite being present in the SLE patient’s blood; this supports the ANA profile enzyme dot immunoassay as a more sensitive test.

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ANA-specific antibodies, ANA patterns, anti-ds-DNA results, and clinical diagnosis: a laboratory and clinical audit

Cited by 3 publications

References 25 publications

Synbiotic regulates gut microbiota in patients with lupus nephritis: an analysis using metagenomic and metabolome sequencing

Synbiotic regulates gut microbiota in patients with lupus nephritis: an analysis using metagenomic and metabolome sequencing

Discriminating between Homogeneous (AC-1) and Dense Fine Speckled (AC-2) Antinuclear Antibody Patterns: Re-Evaluation of Immunofluorescence Imaging

Negative ANA-IIF in SLE patients: what is beyond?

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