Antinuclear Antibody Test Research Articles

Outcomes of first assisted reproductive technology treatment in infertile women with and without antinuclear antibodies: A prospective cohort study

BackgroundIt has been shown that antinuclear antibodies (ANAs) are associated with adverse reproductive events. The presence of ANAs may reduce the pregnancy rate in women undergoing assisted reproductive technology (ART) treatment. MethodsThis study aimed to investigate the potential effect of ANAs on the outcomes of infertile women undergoing in-vitro fertilization and embryo transfer (IVF-ET) or intracytoplasmic sperm injection (ICSI) for the first time. In total, 907 women were enrolled, with 192 (21.14 %) cases in the ANA+ group and 715 (78.86 %) cases in the ANA- group. Baseline data were collected, and associations between ANAs and oocyte maturation rate, high-quality embryo rate, clinical pregnancy rate, live birth rate and miscarriage rate were analysed. ResultsCompared with the ANA- group, the ANA+ group had a considerably lower oocyte maturation rate (0.69 ± 0.31 vs 0.77 ± 0.26; P = 0.003) and a lower high-quality embryo rate (0.53 ± 0.30 vs 0.78 ± 0.92; P < 0.01). ANAs had no effect on clinical pregnancy rate, live birth rate and miscarriage rate in infertile women receiving ART for the first time. ConclusionsThese findings suggest that ANAs reduce oocyte maturation rate and high-quality embryo rate in infertile women undergoing IVF-ET/ICSI for the first time, but do not have a significant effect on clinical pregnancy rate, live birth rate and miscarriage rate. When testing for ANAs is part of the planned treatment protocol, this should be undertaken in advance of starting IVF-ET/ICSI.

A Multi-task learning U-Net model for end-to-end HEp-2 cell image analysis

Antinuclear Antibody (ANA) testing is pivotal to help diagnose patients with a suspected autoimmune disease. The Indirect Immunofluorescence (IIF) microscopy performed with human epithelial type 2 (HEp-2) cells as the substrate is the reference method for ANA screening. It allows for the detection of antibodies binding to specific intracellular targets, resulting in various staining patterns that should be identified for diagnosis purposes. In recent years, there has been an increasing interest in devising deep learning methods for automated cell segmentation and classification of staining patterns, as well as for other tasks related to this diagnostic technique (such as intensity classification). However, little attention has been devoted to architectures aimed at simultaneously managing multiple interrelated tasks, via a shared representation.In this paper, we propose a deep neural network model that extends U-Net in a Multi-Task Learning (MTL) fashion, thus offering an end-to-end approach to tackle three fundamental tasks of the diagnostic procedure, i.e., HEp-2 cell specimen intensity classification, specimen segmentation, and pattern classification. The experiments were conducted on one of the largest publicly available datasets of HEp-2 images. The results showed that the proposed approach significantly outperformed the competing state-of-the-art methods for all the considered tasks.

Genetic Analysis of Asymptomatic Antinuclear Antibody Production.

Antinuclear antibodies (ANA) are detected in up to 14% of the population, and many individuals with ANA are asymptomatic. The literature on the genetic contribution to asymptomatic ANA positivity is limited. In this study, we aimed to perform a genome-wide association study of asymptomatic ANA positivity in multiple populations. Asymptomatic individuals who were either ANA positive or ANA negative from the All of Us Research Program were included in this study, selecting those with an ANA test performed by immunofluorescence and no evidence of autoimmune disease. Imputation was performed, and a multipopulation meta-analysis including approximately 6 million single-nucleotide polymorphisms (SNPs) was conducted. Genome-wide SNP-based heritability was estimated using the Genome-wide Complex Trait Analysissoftware. A cumulative genetic risk score for lupus was constructed using previously reported genome-wide significant loci. A total of 1,955 asymptomatic ANA positive and 3,634 asymptomatic ANA negative individuals across three populations were included. The multipopulation meta-analysis revealed SNPs with a suggestive association (P <1 × 10-5) across 8 different loci, but no genome-wide significant loci were identified. A gene variant upstream of HLA-DQB1, (rs17211748, P = 1.4 × 10-6, odds ratio 0.82, 95% confidence interval 0.76-0.89), showed the most significant association. The heritability of asymptomatic ANA positivity was estimated to be 24.9%. Individuals who were asymptomatic and ANA positive did not exhibit increased cumulative genetic risk for lupus compared with individuals who were ANA negative. ANA production is not associated with significant genetic risk and is primarily determined by environmental factors.

Successful treatment of aspergillosis in a 63-years old female patient with a SIOOT protocol of oxygen-ozone autohemotherapy: A case report

BackgroundInfections caused by the Aspergillus genus are relatively uncommon and typically associated with hospitalization, while only a few cases are linked to environmental exposure to filamentous moulds. These infections can significantly impact pulmonary function, and antifungal therapy may further compromise lung physiology, especially if fungal nodules develop. Consequently, many cases of aspergillosis are not directly treated to avoid additional lung damage. In this case report, we explore a novel approach using ozone therapy to improve clinical outcomes in a patient with pulmonary aspergillosis. To our knowledge, no previous data on the use of this procedure in treating Aspergillus infections have been published in the scientific literature. Case presentationA 63-year-old Caucasian female physician presented with persistent cough, mild fever, mucous sputum, fatigue, and general discomfort. After extensive laboratory analyses, she was initially suspected of having a Mycobacterium tuberculosis infection but was later accurately diagnosed with Aspergillus fumigatus. Allergy tests, thyroid function assessments, total-body PET, and maxillofacial imaging did not yield any findings to suggest alternative diagnoses. Aside from mild neutropenia and a positive antinuclear antibody (ANA) test, which appeared to be laboratory artifacts, the patient showed no significant inflammatory biomarkers, thyroid dysfunction, or signs of allergy. Chest CT revealed Aspergillus nodules with only mild obstructive respiratory impairment, which did not worsen upon administration of salbutamol. Following a period of mild symptoms, her health declined in the summer of 2022, prompting her to pursue ozone therapy at a colleague’s recommendation. After twelve oxygen-ozone treatments, her symptoms—including mild fever, cough, discomfort, and fatigue—fully resolved, and a follow-up chest CT showed a reduction in the presence of Aspergillus. ConclusionOxygen-ozone therapy administered through autohemotherapy proved to be a promising treatment for nodular aspergillosis in the lung, highlighting the need for further exploration of the mechanisms behind this positive outcome. This report is particularly relevant for physicians trained in ozone therapy, as it requires specialized professional expertise. This is the first documented case showing a successful outcome of ozone therapy in treating nodular Aspergillus fumigatus lung infection with mild obstructive pulmonary impairment. The favourable results should encourage physicians to consider this therapeutic approach in the future.

Automatic classification of HEp-2 specimens by explainable deep learning and Jensen-Shannon reliability index

The Anti-Nuclear Antibodies (ANA) test using Human Epithelial type 2 (HEp-2) cells in the Indirect Immuno-Fluorescence (IIF) assay protocol is considered the gold standard for detecting Connective Tissue Diseases. Computer-assisted systems for HEp-2 image analysis represent a growing field that harnesses the potential offered by novel machine learning techniques to address the classification of HEp-2 images and ANA patterns.In this study, we introduce an innovative platform based on transfer learning with pre-trained deep learning models. This platform combines the power of unsupervised deep description of HEp-2 images, a novel feature selection approach designed for unbalanced datasets, and independent testing using two distinct datasets from different hospitals to tackle cross-hardware compatibility issues. To enhance the trustworthiness of our method, we also present a modified version of gradient-weighted class activation mapping for regional explainability and introduce a new sample quality index based on the Jensen-Shannon divergence to enhance method reliability and quantify sample heterogeneity.The results we provide demonstrate exceptionally high performance in intensity and ANA pattern recognition when compared to state-of-the-art approaches. Our method's ability to eliminate the need for cell segmentation in favor of statistical analysis of the sample makes it applicable, robust, and versatile. Our future work will focus on addressing the challenge of mitotic spindle recognition by expanding our proposed approach to cover mixed patterns.

HLA-DRB1*01 predicts treatment outcome in juvenile idiopathic arthritis: A retrospective-prospective cohort study.

Juvenile idiopathic arthritis (JIA) is the most common chronic inflammatory autoimmune disease in childhood, significantly contributing to both short- and long-term disability. While certain human leukocyte antigen (HLA) class II alleles are known to be associated with specific subgroups of JIA, emerging evidence suggests a strong correlation between these alleles and treatment response. This study involved 143 JIA patients diagnosed according to International League of Associations for Rheumatology criteria. Each patient underwent HLA class II typing, including HLA-B27, as well as tests for rheumatoid factor (RF) and antinuclear antibodies (ANA). Comprehensive rheumatological assessments were conducted at diagnosis, with follow-ups at three and six months post-onset. After six months of methotrexate (MTX) treatment, patients were categorized as responders or non-responders. Responders achieved clinically inactive disease based on the American College of Rheumatology Provisional Criteria for Defining Clinical Inactive Disease and Clinical Remission. Non-responders, who did not reach clinically inactive disease after six months of treatment, required the addition of another non-biological disease-modifying antirheumatic drug (DMARD) or a biological DMARD. Our analysis revealed that the HLA-DRB1*01 allele is a significant prognostic marker for therapeutic response, predicting therapeutic resistance (P=0.01). The most prevalent HLA-DRB1 alleles in the treatment-resistant group were HLA-DRB1*08:11 (11.3%), HLA-DRB1*01:01 (8.5%), HLA-DRB1*01:13, HLA-DRB1*04:11 (7%), HLA-DRB1*08:13, and HLA-DRB1*08:15 (4.2%). These findings highlight the critical role of HLA class II alleles in pediatric rheumatology, particularly in relation to treatment response and disease prognosis. In the era of personalized medicine, understanding the genetic contributions to treatment response and outcomes in JIA patients is essential. A key limitation of this study was the lack of comparison of treatment responses across different JIA subtypes. Future studies should prioritize evaluating MTX efficacy within specific JIA subgroups to enable a more tailored understanding of its effectiveness.

Mitigating overuse of antinuclear antibody (ANA) testing through educational intervention: a study in internal medicine and neurology departments

Introduction/objectivesOveruse of antinuclear antibody (ANA) tests leads to increased costs, false positives, and unnecessary treatments. This study evaluated ANA overuse in internal medicine and neurology departments and assessed the impact of an educational intervention.MethodThis quality improvement educational intervention study examined ANA test overuse in five internal medicine departments and one neurology department at a university-affiliated medical center. The educational intervention included a session focusing on ANA testing appropriateness. Outcome measures comprised the ANA/new patient ratio (APR) and the percentage of positive ANA test results. Outcomes were compared between the pre- and post-intervention periods (both 6 months).ResultsThe intervention took place in December 2021. The APR decreased from 43% in the pre-educational intervention period to 27% in the post-intervention period in the neurology department (odds ratio [OR] 0.49, confidence interval [95% CI] 0.37–0.63, P < 0.0001) and from 2.6% to 2.2% in the internal medicine departments (OR 0.89, 95% CI 0.73–1.10, P = 0.28). The percentage of positive ANA tests increased from 43% pre-intervention to 53% in the post-intervention period (OR 1.49, 95% CI 0.90–2.46, P = 0.12) in the neurology department and from 48% to 59% (OR 1.56, 95% CI 0.99–2.44, P = 0.0543) in the internal medicine departments.ConclusionA simple educational intervention reduced unnecessary ANA testing in the neurology department but not in internal medicine departments, improving patient selection and potential cost savings. The results underscore the importance of targeted education to promote evidence-based behavior among healthcare professionals. Further research with longer follow-up is needed to assess the sustainability of these improvements.Key Points• This study presents a quality improvement intervention to prevent the overuse of ANA testing in non-rheumatologist physicians.• A straightforward educational intervention effectively reduced unnecessary ANA testing and improved patient selection, following the Choosing Wisely campaign.• This educational intervention can be adapted and applied in various clinical settings to address the overuse of diagnostic tests, benefiting a wide range of healthcare professionals.

Challenges in Diagnosis and Management of SLE in Africa: An Online Survey.

We surveyed African physicians about challenges in diagnosis and management of systemic lupus erythematosus (SLE). We used a cross-sectional, online questionnaire-based survey of African specialist physicians on availability of laboratory tests, medications, and specialized services for the diagnosis of SLE. Our 226 respondents from 31 countries were dermatologists (38%), rheumatologists (28%), and nephrologists (23%), the majority practicing at university/state-funded hospitals (80.8%), but over half of patients (59.6%) were self-funded for laboratory tests and medications. Antinuclear antibody (ANA), antiphospholipid antibody, and complement tests were available to 79.4%, 67.6%, and 62.3% of respondents, respectively, but fewer in the East and West African regions. Median turnaround time for the ANA test was within two weeks but more than four weeks for 5.6% of respondents, and longer in West Africa compared with other regions (P = 0.0002). Availability of urine protein-to-creatinine test, skin and renal histopathology was 82%, 82.5%, and 76.2%, respectively. Median turnaround times were within one to two weeks, but more than four weeks for 13.8% of respondents for skin histology results and usually within four weeks but more than four weeks for 24.5% of respondents for renal histology. Glucocorticoids and antimalarials were readily available across all regions, with variable availability of immunosuppressants from 93.7% for methotrexate to 65% for calcineurin inhibitors and only 58.4% for the biologic rituximab. Intensive care units/high care facilities, hemodialysis, and renal transplantation were available to 69.8%, 91.9%, and 56.5% of respondents, respectively. Variable availability of laboratory tests, medications, and supportive services coupled with cost constraints are major impediments to early diagnosis and optimal management of SLE in most of Africa and are likely factors contributing to underreporting and poor prognosis of SLE in Africa.

B-082 Clinical Investigation of Rods and Rings Pattern in Anti-Nuclear Antibody Test in Korean Patients

Abstract Background In the mid-2000s, a novel pattern in antinuclear antibody (ANA) testing was reported. Indirect immunofluorescence assays on HEp-2 cells, conducted with sera from patients with Hepatitis C virus (HCV) infection, revealed cytoplasmic structures in the form of rods and rings (RR). These structures typically displayed rod lengths of 3-10μm and ring diameters of 2-5μm. Inosine monophosphate dehydrogenase (IMPDH) was identified as the primary component of the RR structures. Research on the administration of interferon/ribavirin (IFN/RBV) showed that ribavirin, an IMPDH inhibitor, induced RR formation in over 95% of cells. Additionally, RR patterns were reported in patients receiving other IMPDH inhibitors such as mycophenolic acid and azathioprine. While cytidine triphosphate synthase 1 (CTPS1) was suggested to be associated with RR structures, there were no reported cases in patients. Interestingly, patients receiving methotrexate, which does not directly inhibit IMPDH but hinders GTP biosynthesis, also exhibited RR structures. To explore the clinical implications of specimens showing RR patterns, a study was conducted to investigate the actual clinical scenarios associated with these findings. Methods This is a retrospective study conducted at a single center, covering the period from January 2022 to December 2023, focusing on the results of ANA testing. The ANA tests were performed using HEp-2 cells (FLUORO HEPANA TEST, MBL, Nagoya, Japan) on slides manufactured with AP 16 IF Plus (das, Palombara Sabina, RM, Italy). ANA results were obtained through qualitative testing, ranging from 1+ to 4+, or quantitative testing based on dilutions ranging from 1:40 to 1:640, depending on clinical requirements. Results Out of the 7,648 ANA test results, the RR pattern was observed in 40 cases (0.5%). Among these, 2.5% had a history of HCV infection with IFN/RBV treatment, and an additional 2.5% had concurrent use of IMPDH inhibitors. There were no reported cases of medication history related to the synthetic pathway of nucleic acids, including CTPS1. Cases where RR patterns coexisted with autoimmune diseases accounted for 35%, infections for 25%, and other inflammatory conditions for 22.5%. RR patterns reported with an intensity of 3+ or at dilutions of 1:160 or higher constituted 27.5%. Among cases displaying strong fluorescence, 45% were associated with infections, 18% with autoimmune conditions, and another 18% with unexplained inflammatory causes. In the overall sample, elevated CRP or ESR was noted in 54.5% of cases with infections, autoimmune diseases, or other inflammatory conditions. Among samples with intense intensity, 55.5% showed elevated CRP or ESR when inflammatory conditions were present. Conclusions The majority of cases exhibiting the RR pattern showed no history of exposure to IMPDH inhibitors or drugs affecting the nucleic acid synthetic pathway. A high proportion of cases clinically diagnosed with inflammation showed a concurrent RR pattern, with only approximately half of them demonstrating elevated CRP or ESR levels. This study suggests the possibility of identifying inflammatory responses not captured by conventional acute inflammation markers through the RR pattern observed in ANA testing.

SARS-CoV-2 induced vitamin D deficiency and psychological stress: a manifestation of autoimmune disease onset.

The global SARS-CoV-2 pandemic significantly altered lifestyles, access to healthcare, and social interactions, introducing unprecedented physical and psychological stress all over the world. This study explores the relationship between psychological stress, vitamin D (Vit-D) levels, and autoimmune connective tissue diseases, including systemic lupus erythematosus, systemic sclerosis, polymyositis, dermatomyositis, and rheumatoid arthritis. The analysis was based on over one million patient data points derived from anti-nuclear antibody (ANA) testing conducted both before and during the COVID-19 pandemic 2017-2021. In a subset of patients, longitudinal data were collected bi-yearly to yearly over 5-8 years using the same three-month criteria. The dataset was analyzed using GraphPad Prism9 using paired t-tests or ordinary one-way ANOVA with a significance threshold of p < 0.05 to ensure robust correlations between the variables. Data indicated that Vit-D levels peaked between 2017 and 2019 before declining, while ANA data demonstrated a rise in autoimmune connective tissue disease cases during the pandemic, reaching a peak in 2021. A clear correlation was observed, with autoimmune disease incidence increasing as Vit-D levels decreased. In-depth case analysis revealed that declining Vit-D levels preceded higher ANA titers and increased autoimmune disease severity, whereas improvements in Vit-D levels were associated with reduced ANA titers and less severe disease manifestations. The findings suggest that maintaining mental health and ensuring adequate Vit-D supplementation could be essential strategies for mitigating autoimmune disease risks and maintaining immune stability, particularly in pandemic scenarios. Clinically, these results underscore the need for early interventions targeting both psychological well-being and Vit-D levels to reduce the burden of autoimmune diseases.

Systemic Lupus Erythematosus, An Unusual Presentation: A Case Report In Azal Hospital, Sana’a, Yemen

Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder characterized by various clinical presentations and potential involvement of multiple organ systems; it exhibits a significant female predominance, with a peak incidence of middle-aged individuals. In this article, we report a case of a 19 years old female patient who presented with fever, headache, fatigue and neck stiffness. Initial investigations revealed a normal white blood cell count (WBC) with neutrophilia, leading to a diagnosis of bacterial meningitis. The patient responded to antibiotic and analgesic treatment. However, subsequent development of arthritis and skin lesions raised suspicion of systemic lupus erythematosus, which was confirmed by positive anti-dsDNA and antinuclear antibody (ANA) tests.

Hep-2000 indirect immunofluorescence assay - An emerging screening tool for detection of antinuclear autoantibodies in various connective tissue disorders.

Connective tissue disorders (CTD's) are a group of autoimmune disorders having multifactorial etiology, multisystem involvement and overlapping clinical features. Their prevalence has been increasing in India, with Systemic lupus erythematosus (SLE) being the most common CTD, affecting mostly females. Antinuclear Antibodies (ANA) directed against a variety of nuclear antigens detectable in the serum are used for screening, diagnoses, and monitoring of autoimmune diseases, with immunofluorescence assay (IFA) and enzyme-linked immunosorbent assay (ELISA) being the most widely used methods. 1.To evaluate the diagnostic significance of IFA in screening of Autoimmune CTDs. 2.To study different titres and patterns shown by ANA positive samples. For IFA, patient's sera is incubated with substrate cells, and bound antibodies are detected by incubation with a dye-conjugated anti-human immunoglobulin which are visualized by fluorescent microscopy and different ANA titres and patterns were analysed. 57 samples were examined for ANA by Indirect IFA, of which 21 (36.8%) were ANA positive with a female preponderance (71.9%) in the peri-menopausal age group. Most common pattern reported was Speckled followed by Homogenous. The sensitivity & specificity of IIFA was found to be comparable with ELISA. CTD's are a group of autoimmune disorders with a plethora of clinical presentations, necessitating the need of a more specific and accurate screening test. ANA by IIFA gives patterns, which are associated with specific antibodies that help in reaching a diagnosis. ANA testing is a cost effective and non-invasive technique that can be used as a reliable screening test for Autoimmune disorders.

The Relation of Neutrophil Lymphocyte Ratio to the Steatosis and Fibrosis Stage in Patient with Non-Alcoholic Fatty Liver Disease Using Fibroscan with Controlled Attenuation Parameter and FIB-4 and FIB-5 Scores

Abstract Background Non-alcoholic fatty liver disease (NAFLD) is a chronic inflammatory disease with excessive fat accumulation in the liver. Fibroscan with controlled attenuation parameter (CAP), a noninvasive imaging procedure in diagnosis of NAFLD patients. Also Neutrophil / lymphocyte ratio (NLR) is a simple marker for inflammation, may be used to predict disease outcome. Objectives Is to determine whether there is a correlation between NLR and the degree of steatosis and fibrosis measured by fibroscan with CAP, FIB-4 and FIB-5 score. Patients and Methods This was a cross sectional study involving 2 groups, group (1): included 115 patients of NAFLD and group (2): included 30 healthy controls. All participants were supjected to complete personal and medical history (history of other comorbidities as DM, Hypertension syndrome and drug/herb intake). Complete physical examination including anthropometric measurements, vital signs, local examination and general examination of any signs of liver dysfunction as hepatomegaly, Jaundice, ascites, splenomegaly and varices. Laboratory tests including (alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), alkaline phosphatase (ALP), Bilirubin, hemoglobin A1c, fasting glucose level, Lipid profile, Antinuclear antibody (ANA) test (to exclude autoimmune hepatitis), serum Uric acid and viral markers (HBsAg &amp; Hcv Ab), Abdominal ultrasound, Fibroscan with CAP and FIB-4 and FIB -5 scoring, complete blood count with calculation of neutrophils /lymphocytes ratio(NLR). Results Both studied groups were comparable in age and sex distribution. Out of NAFLD cases (group 1), more than half (53.9%) had comorbidities (manifested as DM in 37.4%, hypertension in 36.5%, CVD in 8.7% and hypothyroidism in 1.7% of patients), there was no statistically significant difference between the studied groups regarding anthropometric measurements, NAFLD cases (group 1) had significantly lower Hb, HDL level and PLT p value &amp;lt;0.001 and higher NLR, ALT, AST, GGT, cholesterol, triglycerides and LDL levels p &amp;lt; 0.001,there was a significant positive correlation between NLR and each of steatosis and fibrosis p &amp;lt; 0.001,NLR was significantly positively correlated with FIB-4 (r = 0.667, P &amp;lt; 0.001) and FIB-5 (r = 0.361, P &amp;lt; 0.001), NLR was a significant predictor of hepatic steatosis (AUC= 0.816, P = 0.011) and fibrosis (AUC=0.739, P &amp;lt; 0.001) in NAFLD cases. Conclusion The study conducted on NAFLD patients at Ain Shams University Hospital found significant associations between NLR and liver steatosis/fibrosis. NLR proved to be a valuable predictor, with optimal cutoff values for steatosis and fibrosis, aiding in noninvasive assessment and management of NAFLD patients.

Antinuclear antibody staining patterns by indirect immunofluorescence assay observed in patients from a tertiary health center in Latin America

Introduction/ObjectiveThis study aimed to describe the frequency of antinuclear antibody (ANA) staining patterns by indirect immunofluorescence assay observed in patients from a tertiary health center in Latin America. Materials and methodsThis retrospective, descriptive, and observational study evaluated data from all patients undergoing antinuclear antibody indirect immunofluorescence assay from a single-tertiary center (University Hospital Fundación Valle del Lili, Cali-Colombia) in 2020. ResultsOne thousand and eight patients met the inclusion criteria. The median patient age was 47 (34–59.2) years, and most were female (769, 75.3%). A positive ANA immunofluorescence assay was observed in approximately two-thirds of patients (664, 65.8%). ANA test results were primarily used to exclude a suspected diagnosis in approximately half of the patients (466, 46.2%). Thirty-seven percent (250/664) of the cohort with ANA-positive titers had a systemic autoimmune rheumatic disease (SARD). The most prevalent SARDs included rheumatoid arthritis (RA) (55, 8.2%) followed by systemic lupus erythematosus (SLE) (37, 5.5%). The vast majority of ANA-positive patients had a reported speckled pattern (anti-cell [AC]-2,4,5; 269; 40.5%) followed by homogenous (AC-1; 266; 40%), nucleolar (AC-8,9,10; 46; 6.9%), and centromere (AC-3; 16; 2.4%). The most frequent patterns observed among SLE patients included homogenous (AC-1) patterns in 17 (45.9%) patients, speckled (AC-2,4,5) nuclear patterns in 11 (29.7%) patients, mixed patterns in 7 (18.9%) patients, and reticular/anti-mitochondrial antibody (AMA, AC-21) cytoplasmic patterns in 2 (5.4%) patients. ConclusionThis study is the first to describe ANA patterns in a Colombian population. Speckled and homogenous patterns were predominant in patients with SARDs.

Identifying heterogeneous subgroups of systemic autoimmune diseases by applying a joint dimension reduction and clustering approach to immunomarkers

BackgroundThe high complexity of systemic autoimmune diseases (SADs) has hindered precise management. This study aims to investigate heterogeneity in SADs.MethodsWe applied a joint cluster analysis, which jointed multiple correspondence analysis and k-means, to immunomarkers and measured the heterogeneity of clusters by examining differences in immunomarkers and clinical manifestations. The electronic health records of patients who received an antinuclear antibody test and were diagnosed with SADs, namely systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and Sjögren’s syndrome (SS), were retrieved between 2001 and 2016 from hospitals in Taiwan.ResultsWith distinctive patterns of immunomarkers, a total of 11,923 patients with the three SADs were grouped into six clusters. None of the clusters was composed only of a single SAD, and these clusters demonstrated considerable differences in clinical manifestation. Both patients with SLE and SS had a more dispersed distribution in the six clusters. Among patients with SLE, the occurrence of renal compromise was higher in Clusters 3 and 6 (52% and 51%) than in the other clusters (p < 0.001). Cluster 3 also had a high proportion of patients with discoid lupus (60%) than did Cluster 6 (39%; p < 0.001). Patients with SS in Cluster 3 were the most distinctive because of the high occurrence of immunity disorders (63%) and other and unspecified benign neoplasm (58%) with statistical significance compared with the other clusters (all p < 0.05).ConclusionsThe immunomarker-driven clustering method could recognise more clinically relevant subgroups of the SADs and would provide a more precise diagnosis basis.

Detection and differentiation of antinuclear antibodies in serum of dengue suspected patients with or without systemic autoimmune disease in Kolkata, India

ABSTRACT The pathophysiology of dengue may be influenced by antibodies released during infection. Several autoimmune diseases are accompanied by antinuclear antibodies (ANAs) but 8–10% of the general population have positive ANA tests. To test the hypothesis that an ANA-positive test indicates an immune dysregulated state that modifies the risk for certain clinical disorders in people with or without an autoimmune disease, we examined the various ANA profiles and their relationships to various autoimmune disorders, as well as the severity of these relationships, in patients infected with dengue fever. Enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) methods were used. Indirect immunofluorescence assay (IIFA) and line immunoassay (LIA) were performed to detect and differentiate the ANAs among dengue infected patients. Out of 135 dengue virus-positive patients, 94.07% were positive by ELISA and 5.93% positive by RT-PCR method. ANAs by IIFA and LIA were detected in 54.8% and 18.5% of the dengue positive patients, respectively, and 10.3% and 7.1% of the 126 dengue negative patients, respectively. This study showed that dengue was associated with an increased risk of autoimmune myositis and mixed connective tissue disease (MCTD), a rare complication of dengue. The risk of other autoimmune diseases did not seem to increase after DENV infection.

Antinuclear Antibodies in Systemic Lupus Erythematous – Their Complex Role and Clinical Significance in Diagnosis

Antinuclear antibodies (ANA) are a wide group of proteins directed against autologous cellular components, primarily nucleic acids and histones. Their levels are assessed using immunofluorescence on Hep-2 cells or a solid-phase ANA screening immunoassay to subsequently obtain titer value with positive cut-point of ≥1:80. Studies show that ANA can be found in 13% of general population, but typically they are associated with autoimmune conditions and inflammatory connective tissue diseases for example: systemic lupus erythematosus (SLE), sjögren's syndrome, rheumatoid arthritis, mixed connective tissue disease, juvenile idiopathic arthritis, systemic sclerosis, inflammatory myopathies. ANA detection is especially important for SLE, where 2 antibody types - anti-Sm and anti-dsDNA - serve as an entry diagnostic criterion. With increasing patient screening for those antibodies, it is important to determine that alone, positive ANA assays cannot confirm nor deny any disease and to classify their presence as a marker of a disease it is required to satisfy additional additive criteria approved in 2019 by European League Against Rheumatism/American College of Rheumatology. SLE diagnosis can be made when patient collects 10 points from clinical or immunologic domains described by EULAR/ACR, which makes SLE diagnosis challenging and therefore, describing the guidelines is vital to remain cautious about overestimation of the position positive ANA values hold in clinical practice.In conclusion the positive ANA test may be a basis for diagnosis, when additional symptoms occur, but alone does not hold any diagnostic significance and may lead to unnecessary stress for patient.

The Diagnostic Role of Antinuclear Antibody: A Study of Clinical Utility in a Tertiary Hospital

Abstract Introduction: Various studies reported unnecessary and inappropriate serology testing of rheumatic diseases leading to a significant waste in healthcare utilization. The purpose of this study is to evaluate the clinical utility of antinuclear antibody (ANA) testing in a real-world setting within a tertiary hospital for systemic lupus erythematosus (SLE) and other ANA-associated rheumatic diseases (AARDs), as well as to identify patient characteristics and test results that predict rheumatic disease association. Subjects and Methods: This is a retrospective study of patients aged 15 years or older who underwent ANA testing at Aseer Central Hospital from January 2018 to December 2022. Data collected included patient demographics, clinical presentations, referral physician type, ANA test results, and final diagnoses. Descriptive statistics characterized patient demographics and ANA test results. Sensitivity, specificity, and predictive values of ANA testing were calculated for SLE and AARD diagnoses. Chi-squared test was used to identify the predictive values of AARDs. Results: Of the 2141 patients tested for ANA at Aseer Central Hospital, 583 (27.2%) tested positive, with a higher proportion of females (80.8%). Notably, 85.1% of patients who tested ANA positive were under 55 years old. The highest ANA test-positive proportion was noticed by rheumatologist physician’s referral (67.6%), referral symptoms including joint symptoms (38.3%), mucocutaneous symptoms (19.7%), renal disease symptoms (14.4%), and hematological abnormalities (14.2%). The sensitivity and specificity of ANA for diagnosing SLE were 86.4% and 79.3%, respectively, with a positive predictive value (PPV) of 31.7% and a negative predictive value (NPV) of 98.1%. The sensitivity and specificity of other AARDs were 85.4% and 73.9%, respectively, with a PPV of 6% and an NPV of 99.6%. Significant associated factors with AARDs included younger age (&lt;55 years), female patients, higher ANA titer, rheumatologist referral, and clinical indications such as sicca symptoms, myopathy, mucocutaneous symptoms, and hematological abnormalities (P &lt; 0.001). Conclusion: In this study, ANA testing showed a good sensitivity and NPV in ruling out AARDs; however, its poor specificity and PPV suggest that positive ANA findings should be interpreted cautiously. Younger age, female gender, higher ANA titer, rheumatologist referral, and specific clinical indications were significantly associated with AARDs, suggesting the importance of targeted ANA testing in clinical practice.

The Relative Timing, Outcomes, and Economic Impact of Anti-Nuclear Antibody (ANA) and Extractable Nuclear Antigen (ENA) Laboratory Ordering.

Objective: To determine the rates of simultaneous antinuclear antibodies (ANA) screening and extractable nuclear antigen (ENA) testing that do not follow recommendations.Design, Setting, and Participants: Retrospective cohort study of adult patients (≥18 years) with a HEp-2 ANA or ENA ordered in the Marshfield Clinic Health System.Main Outcome(s) and Measure(s): Counts of patients having simultaneous ANA and ENA laboratory testing or ENA testing without ANA screening. Relevant ENA positivity in ANA negative patients. Secondary measures included relative timing of ANA and ENA ordering, potential cost savings of unnecessary testing, and provider ordering characteristics including specialty and provider type.Results: Of 58,627 cohort patients, 39,155 (66.8%) were women, and the mean (SD) age at first laboratory testing was 48.7 (19.0) years. The negative ANA with positive ENA rate was 2%. Further stratification identified only 23 diagnosed autoimmune connective tissue diseases (AI-CTDs) in this 2%, with a resulting negative ANA with relevant positive ENA rate of 0.37%. Simultaneous ANA and ENA testing occurred in 8.3% of patients, and an ENA only was ordered in 24.2% of patients. The simultaneous or non-sequential ordering of ANA and ENA testing resulted in significant health care costs of $2,293,251.80 over 20,112 unique patients.Conclusions and Relevance: A significant percentage of providers do not follow recommendations to sequentially order ANA and ENA testing on patients with suspected AI-CTDs. Significant saving in health care spending without failure to diagnose AI-CTDs can be achieved if ANA testing is performed first, followed by ENA testing when suspecting AI-CTDs in patients.

Genetic analysis of asymptomatic antinuclear antibody production.

Antinuclear antibodies (ANA) are detected in up to 14% of the population and the majority of individuals with ANA are asymptomatic. The literature on the genetic contribution to asymptomatic ANA positivity in the population is limited. In this study, we aimed to perform a multi-ancestry genome-wide association study (GWAS) of asymptomatic ANA positivity. Asymptomatic ANA positive and negative individuals from the All of Us Research Program were included in this study, selecting those with an ANA test by immunofluorescence and no evidence of autoimmune disease. Imputation was performed and a multi-ancestry meta-analysis including approximately 6 million single-nucleotide polymorphisms (SNPs) was conducted. Genome-wide SNP based heritability was estimated using the GCTA software. A cumulative genetic risk score for lupus was constructed using previously reported genome-wide significant loci. 1,955 asymptomatic ANA positive and 3,634 asymptomatic ANA negative individuals were included across three genetic ancestries. The multi-ancestry meta-analysis revealed SNPs with a suggestive association (p-value < 1×10 -5 ) across 8 different loci, but no genome-wide significant loci were identified. A gene variant upstream of HLA-DQB1 (rs17211748, P = 1.4×10 -6 , OR = 0.82, 95% CI 0.76-0.89) showed the most significant association. The heritability of asymptomatic ANA positivity was estimated to be 24.9%. Asymptomatic ANA positive individuals did not exhibit increased cumulative genetic risk for lupus compared to ANA negative individuals. ANA production is not associated with significant genetic risk and is primarily determined by non-genetic, likely environmental, factors.