Antinuclear Antibody Test Research Articles

Prediction of a Positive ANA Result for a Rheumatological Diagnosis in an Outpatient Setting

Objective: To study the predictive value of a positive anti-nuclear antibody (ANA) for a rheumatological diagnosis in an outpatient setting. Methods: Individuals who were referred to the rheumatology outpatient clinics because of a positive ANA between July 2014 and June 2015 were retrospectively reviewed. Presenting symptoms in addition to a positive ANA and whether a final rheumatological diagnosis was made were recorded. The positive predictive value of a positive ANA and its titer for a rheumatological diagnosis, with and without accompanying symptoms was evaluated. Results: A total of 230 patients were included (82% women, age 47.7 ± 14.1 years [range 18-84]). Family medicine and the general outpatient clinic were the main sources of referral (32.2%), followed by ophthalmology (13.0%) and otorhinolaryngology (11.7%). A final rheumatological diagnosis was made in 54 (23.5%) patients, with rheumatoid arthritis being the commonest diagnosis (40.7%). In the absence of any associated symptoms, the predictive value of a positive ANA was 0%. The presence of Raynaud’s phenomenon (100%), joint swelling (59.5%), and joint stiffness (48.9%) predicted a better final rheumatological diagnosis along with a positive ANA. ANA titers of 1:80 or less had a low sensitivity for rheumatic diseases. A receiver operating characteristic (ROC) curve analysis showed that an ANA titer of [Formula: see text]1:128 best predicted a rheumatological diagnosis (AUC 0.78 [0.71–0.85]; sensitivity 0.78; specificity 0.64). Conclusions: To improve the prediction for a rheumatological diagnosis, referral for a positive ANA test should be more appropriately done with compatible symptoms.

Case Report: Tuberculosis lymphadenitis with systemic lupus erythematosus in a young woman: a case report

Background: Tuberculosis is a chronic infectious disease and can be categorised into pulmonary TB and extra-pulmonary TB based on its spread. TB lymphadenitis is one of the extra-pulmonary TB diseases. Patients with a weakened immune system in systemic lupus erythematosus (SLE) have an increased incidence of TB. Case: Here we present a case report of a 21-year-old female patient with SLE diagnosed with tuberculous lymphadenitis at dr. Zainoel Abidin Hospital, Banda Aceh, Indonesia. The patient complained of a lump in the right neck 4 months ago with a diameter of 4 cm, accompanied by fever, decreased appetite, and weight loss. Other than that, the patient also experiences joint pain, hair loss and sun sensitivity since 12 months ago. Chest radiography showed no abnormalities, and fine-needle aspiration biopsy results confirmed tuberculous lymphadenitis. Antinuclear antibody test was borderline. The patient had been taking steroids and hydroxychloroquine for the past 10 months. Currently, the patient is taking the advanced phase of antituberculosis drugs FDC. After undergoing the intensive phase of antituberculosis drugs, the submandibular lump got smaller to a diameter of 2 cm. Conclusion: TB lymphadenitis is a rare case but can occur in conditions of decreased immunity like SLE. It involves some of the immune disorders caused by the long-term use of immunosuppressive therapy.

P-172 Effect of anti-centromere antibodies on multi pronuclear formation

Abstract Study question Do the anti-centromere antibodies (ACA) affect the multi pronuclear formation (MPF) rate? Summary answer MPF rate was higher in patients with ACA. The MPF rate of ACA patients with antibody titers ≥160-fold was significantly higher than in titers 40-fold. What is known already ACA is an anti-nuclear antibody (ANA), and specifically recognizes the centromere. Recently, several studies have reported that the rate of oocyte maturation is lower in patients with ACA. Moreover, in MI oocytes collected from ACA patients, female chromosomes were frequently dispersed in the cytoplasm. Study design, size, duration A total of 4756 patients from our clinic were tested for ANA before oocyte retrieval from January 2014 to December 2021. After retrospective ANA testing, patients were classified according to 3 groups; 62 ACA patients (with only ACA), 1134 non-ACA patients (with ANA but not with included ACA) and 3560 non-ANA patients (without ACA and any ANA).We considered ACA-positive levels at titers ≥40. ACA patients were further classified into 6 groups by antibody titer. Participants/materials, setting, methods The MPF rate after ICSI was compared in the 3 groups (ACA patients, non-ACA patients, non-ANA patients) and at 6 ACA titers in each group (titer of ACA; 40-fold, 80-fold, 160-fold, 320-fold, 640-fold, ≥1280-fold).MPF rate was calculated by dividing the number of embryos that formed three or more pronuclei by the number of embryos inseminated by ICSI.Ryan's method was used for multiple comparisons of ratios. Main results and the role of chance MPF rate was 3.8% (1997/53240) in non-ANA patients, 4.3% (733/17003) in non-ACA patients, and 32.1% (351/1092) in ACA patients, being significantly higher in ACA vs other groups (P<.01). In comparisons between ACA titers, MPN rate was 8.7% (4/46) at 40-fold , 13.0% (3/23) at 80-fold , 36.1% (56/155) at 160-fold , 32.4% (48/148) at 320-fold , 36.0% (111/308) at 640-fold , and 31.3% (129/412) at ≥ 1280-fold, respectively. MPN rate of patients with titers ≥160-fold was significantly higher than with 40-fold. Limitations, reasons for caution A potential limitation of the present study is the small sample size. This is because ACA patients account for only 1% of patients who underwent ART treatments. Wider implications of the findings MPF rate in ACA patients was significantly higher than in non-ANA and non-ACA patients and was also significantly higher in ACA patients with titers of ≥ 160-fold vs 40-fold. The dispersion of the female chromosome in the cytoplasm of MI oocytes may be a cause of MPN formation. Trial registration number None

The significance of dense fine speckled pattern in antinuclear antibody-associated rheumatic disease and coexisting autoantibodies: A propensity score-matched cohort study.

To investigate the relationship between the prevalence of antinuclear antibody (ANA) -associated rheumatic diseases (AARD) and the presence of dense fine speckled (DFS) and homogeneous patterns in ANA tests. This retrospective study enrolled adult patients with either a DFS or homogeneous pattern in their ANA test. A mixed pattern was defined as the presence of more than one pattern reported in the test. The presence of anti-DFS70 antibodies and other common autoantibodies were detected using EUROLINE ANA Profile 23. A 1:2 propensity score matching was applied to control for demographic and other interfering factors. A total of 59 patients with a DFS pattern were enrolled and compared with a matched homogeneous group. The DFS group had a significantly lower prevalence of AARD (3.4% vs. 16.9%, p = .008) and the subgroup with anti-DFS70 antibodies showed an even lower prevalence (2% vs. 20%, p = .002). Among the 33 patients with monospecific anti-DFS70 antibodies, five had a mixed pattern, and all patients with common autoantibodies had an isolated DFS pattern. The findings of this study suggest that patients with a DFS pattern in their ANA test may have a lower prevalence of AARD compared with those with a homogeneous pattern. However, an isolated DFS pattern in ANA testing does not necessarily indicate the presence of monospecific anti-DFS70 antibodies or AARD. Confirmatory testing for the monospecific anti-DFS70 antibody is mandatory to exclude AARD.

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.

#2909 DOUBLE GLOMERULOPATHY OF THIN BASEMENT MEMBRANE DISEASE AND IGA NEPHROPATHY PRESENTING WITH NEPHROTIC SYNDROME IN AN ADULT WOMAN

Abstract Background and Aims We report a case of a 44-year-old woman with no known comorbid conditions who, eventually was diagnosed to have simultaneous occurrence of IgA nephropathy and thin basement membrane disease atypically presenting with heavy proteinuria, hypoalbuminemia, bipedal edema, ascites, bilateral pleural effusion, and hemodynamic instability, but with a surprisingly normal light microscopic findings and an Oxford MEST C score of 0 on renal biopsy. who, clinically improved with immunosuppression and angiotensin II receptor blocker therapy. Physical exam revealed an overweight female, with facial swelling, no rashes, periorbital and a Grade II bipedal pitting oedema. The presence of decreased breath sounds and dullness on percussion on both lower lung fields were also observed. Shifting dullness and fluid wave on abdominal exam. The patient was afebrile, no desaturation at room air and a palpatory blood pressure of 70 mmHg. Secondary causes of glomerulonephritis such as Hepatitis B and Hepatitis C infection were ruled out. Antinuclear antibody (ANA) test, compliment (C3 and C4) with anti-dsDNA determination to rule out SLE were unremarkable as well. Serum creatinine was elevated at 2.63 with estimated GFR of 22 mL/min. Further work-up with 24-hour total urine protein was 22,026.2 grams/day The patient underwent percutaneous kidney biopsy which showed IgA Nephropathy with a MEST C score of 0 with simultaneous occurrence of thin basement membrane disease. Method/ Treatment Pulse steroid therapy was started with Methylprednisone 1-gram intravenous infusion once daily for 3 days followed with Prednisone (1mg/kg/day) 60 mg a day and a low-dose of Angiotensin receptor blocker, Candesartan, 4mg once daily for the proteinuria. Results After 8 weeks of corticosteroid and ARB therapy, patient was seen at the clinic with resolution of bipedal edema, no demonstrable ascites and clear breath sounds on chest and lungs auscultation. Blood pressure was 130/80 with no proteinuria on dipstick, random urine protein: creatinine ratio (UPCR) of 0.07, creatinine of 0.97 mg/dl and BUN of 17 mg/dl. Prednisone was then reduced with the goal to taper the dose for a period of 6 months. Conclusion/ Significance Both IgA nephropathy and thin basement membrane disease usually presents with hematuria, hypertension, and varying degrees of proteinuria. However, nephrotic syndrome as its presentation has not been well characterized to date, and whether this adds to the mortality is not yet established. Although the most widely accepted system, the Oxford MEST-C scores is utilized in predicting renal outcomes, its role in double glomerulopathies has not been validated. Even though many investigators have indicated that the presence of heavy proteinuria at the initial evaluations was almost always associated with progressive renal failure and that there is no effective medical treatment aside from supportive care, in this study, a trial of immunosuppression yielded improved clinical outcomes.

Systemic sclerosis with interstitial lung disease and myocardial infarction: a case report.

The authors present a rare case of diffuse SSc with ILD and myocardial infarction having a history of Raynaud phenomenon, skin thickening, and shortness of breath. Antinuclear antibody and antitopoisomerase antibody tests were positive. The patient was managed medically and the condition of patient is improving. SSC can affect the skin as well as other organs, with the lungs being the most frequently involved and seriously impacted. SSc patients can have multiple organ involvement like the skin, lungs, heart, kidneys, and gastrointestinal tract. Because ILD is the leading cause of death among people with SSC, early diagnosis and high suspicion of lung involvement can reduce mortality. The mortality rate for SSC associated with ILD is extremely high. Even though ILD is common in SSc, it might be difficult to identify and detect early for which a high-resolution CT scan can be used. In SSc patients, heart involvement can coexist with ILD.

P196 ENDOCARDITIS MASQUERADING AS STILL‘S DISEASE: WHEN AN ECHOCARDIOGRAM SAVES YOUR LIFE

Abstract A 44–year–old man in apparent good health thus far presented with weight loss and fever preceded by shivers, profuse sweating, and arthralgias. Blood tests documented normal white blood count with slightly altered CRP and ESR. Chest X–rays did not find pleuro– parenchymal lesions with normal cardiac silhouette. Abdomen ultrasounds was regular. Due to the persistence of the symptoms, he was referred to a rheumatologist who diagnosed Adult Still’s disease (AOSD) and recommended therapy with prednisone and sulfasalazine, interleukin–1 inhibitor, with the resolution of the fever. After three months, because of the recrudescence of the fever, he was admitted to the emergency room a total–body CT–scan documented inflammatory–abscess lesions in the brain and hypodense areas in the spleen. ECG showed type 1 second–degree atrioventricular (AV) block. The patient was therefore transferred to the Cardiology Department; echocardiograms documented an endocardic mass on the aortic valve and suspected periannular abscess also infiltrating the interventricular septum. The patient underwent urgent surgery with excision of the aortic valve, opening and draining of the abscessual cavities, reconstruction of the aortic annulus with bovine pericardium patch and aortic valve replacement with bioprosthesis. The patient did not present further febrile episodes after surgery but a pacemaker implantation was necessary due to persistent AV block. Biological material sent for culture was positive for S. gallolyticus. AOSD is one of the most common conditions responsible for fevers of unknown origin. It is characterized by fever, arthritis, evanescent rash, leukocytosis, and multiorgan involvement. Yamaguchi criteria required for the diagnosis of AOSD are as follows: main criteria: Fever >39°C, lasting 1 week or more, Arthralgia or arthritis, lasting 2 weeks or more, Typical rash, Leukocytosis >10,000/mm3; Minor criteria: Sore throat, Recent development of significant lymphadenopathy, Hepatomegaly or splenomegaly, Negative tests for antinuclear antibodies and rheumatoid factor; Exclusion criteria: Infections, Malignancies, Other rheumatic diseases.

Clinical usefulness of anti-nuclear antibody in childhood: real-world experience at a tertiary care center : Usefulness of ANA in pediatric autoimmune diseases.

We evaluated the reasons for requesting anti-nuclear antibody (ANA) analysis in clinical practice at a tertiary center and the performance of ANA in pediatric autoimmune diseases. Patients under 18years of age who underwent ANA testing for various symptoms between 2013 and 2017 were included. We retrieved data from medical records, including demographic and clinical characteristics, diagnoses, ANA results, titers, and staining patterns. The performance assessment tools were calculated according to the ANA titer for autoimmune diseases. Risk factors for autoimmune diseases in ANA-positive patients were evaluated using logistic regression analysis. Changes in ANA titer and seroconversion were evaluated using repeated ANA analyses. A total of 3812 patients underwent ANA. Medical records of 3320 patients were obtained. The rate of ANA positivity was 27.4%. ANA was requested most frequently because of musculoskeletal findings in 1355 patients (40.8%). Juvenile idiopathic arthritis (n = 174, 20.2%) was the most common diagnosis in ANA-positive patients, followed by systemic lupus erythematosus (n = 52, 6%). For autoimmune diseases, a titer of ≥ 1:100, a sensitivity of 40.1%, and a specificity of 77.1% were observed. At a titer ≥ 1:1000, the sensitivity and specificity were 24.1% and 89%, respectively. Homogeneous staining was an additional risk factor for autoimmune diseases in ANA-positive patients by multivariate logistic regression analysis (OR [95% CI]: 4.562 [3.076-6.766], p < 0.001). Conclusion: Our results revealed that the performance of the ANA test in diagnosing autoimmune diseases in pediatric clinical practice was poor. Therefore, clinical findings should be carefully evaluated before ANA testing is performed. What is Known: • ANA can be detected in systemic autoimmune rheumatic diseases. • The diagnostic role of ANA is controversial, especially in childhood. What is New: • One in four patients who requested the ANA test had an autoimmune disease. • Less than half of patients with an autoimmune disease had ANA positivity.

Antinükleer Antikor Pozitif Hastalarda Test İsteme Nedenleri ve Hastaların Nihai Tanıları

Background: The aim of this study was to determine the reasons for the request for antinuclear antibody (ANA) in ANA-positive patients and to determine the final diagnosis of these patients and whether they developed a rheumatologic disease. Method: In this retrospective study, the files of 559 patients with positive ANA were reviewed. Demographic, laboratory and clinical characteristics of the patients were noted. At the end of follow-up, the final diagnosis was recorded. Results: The study included 346 patients. 233 of the patients were female, and 113 were male. The mean age at the time of ANA positivity was 9.4  4.7 years, and the mean follow-up period was 19  5.7 months. The most common symptom was myalgia/arthralgia (21.7%). Other common reasons were urticaria, abdominal pain, thrombocytopenia, and proteinuria. Extractable nuclear antigens (ENA) panel results were negative in 170 patients (49.1%). In the ENA panel, dense fine speckled antigen 70 antibodies were most frequently positive in 135 patients (39.2%). At the end of follow-up, 234 patients had no disease. One hundred and one patients were diagnosed with non-rheumatologic diseases, and 11 patients were diagnosed with rheumatologic diseases. Eleven patients with rheumatologic diseases were girls. Rash was the most common symptom in patients with rheumatologic diseases. The positive predictive value of ANA positivity for rheumatologic disease was 3.2% and 1.7% for systemic lupus erythematosus. Conclusions: Due to the low positive predictive value of ANA testing, patients at risk for autoimmune diseases should be identified and carefully evaluated before ANA is requested.

A Comparison Between Somatosensory Evoked Potentials and Spine MRI in the Diagnosis of Non-compressive Myelopathy: Which Is More Accurate?

Non-compressive myelopathy is a neurological disorder due to pathological processes affecting the spinal cord in the absence of clinical and radiological evidence of spinal cord compression. Two commonly used diagnostic tools for non-compressive myelopathy are somatosensory evoked potentials (SSEPs) and magnetic resonance imaging (MRI). SSEPs are a neurophysiological tool used to assess the functional integrity of the spinal cord. MRI, on the other hand, is the mainstay imaging modality used for identifying compressive lesions and other structural abnormalities in the spinal cord.The aim of this study was to test the diagnostic accuracy of SSEPs versus spine MRI in the diagnosis and assessment of the severity of non-compressive myelopathy using the Modified Japanese Orthopaedic Association (mJOA) clinical severity score. Our study included 63 subjects. Whole spine MRI and SSEPs (median and tibial SSEP bilaterally) were done for all subjects; their results were compared according to their relation to the mJOAscore and classified into mild, moderate, and severe. The control group was examined to establish normative data for SSEPresults and compared with cases. Blood investigations such as complete blood count, thyroid function test, A1C, HIVtests, venereal disease research laboratory test, erythrocyte sedimentation rate, C-reactive protein, and antinuclear antibodytests were done. Blood tests for vitamin B12 levels were done for patients who were suspected of sub-acute combined degeneration of the spinal cord; cerebrospinal fluid (CSF) analysis was done for patients suspected of multiple sclerosis (MS), acute transverse myelitis (ATM), or other inflammatory/infectious diseases. CSF was analyzed for cell count, cytology, protein, and oligoclonal bands (if indicated). No mild categories were registered in this study; 30%of patients were moderateand 70%were severe. Causes for non-compressive myelopathy in this study werehereditary degenerative ataxias in 12 (38.71%), ATM in 8 (25.81%), and MS in 5 (16.13%); other causes included vitamin B12 deficiency in 2 (6.45%), ischemia in 2 (6.45%), and an unknown cause in 2 (6.45%). SSEPs showed abnormal results in all patients (31; 100%) whereas MRI showed abnormality in only seven patients (22.6%). SSEPsensitivity for detecting severe cases was around 63.6% while that for MRI was 27.3%. The study concluded that SSEPs were more reliable for the detection of non-compressive myelopathies rather than MRI and correlated better with clinical severity. Performing SSEPs is recommended for all patients with non-compressive myelopathy, especially those with negative imaging.

Deep Active Learning for Automatic Mitotic Cell Detection on HEp-2 Specimen Medical Images

Identifying Human Epithelial Type 2 (HEp-2) mitotic cells is a crucial procedure in anti-nuclear antibodies (ANAs) testing, which is the standard protocol for detecting connective tissue diseases (CTD). Due to the low throughput and labor-subjectivity of the ANAs' manual screening test, there is a need to develop a reliable HEp-2 computer-aided diagnosis (CAD) system. The automatic detection of mitotic cells from the microscopic HEp-2 specimen images is an essential step to support the diagnosis process and enhance the throughput of this test. This work proposes a deep active learning (DAL) approach to overcoming the cell labeling challenge. Moreover, deep learning detectors are tailored to automatically identify the mitotic cells directly in the entire microscopic HEp-2 specimen images, avoiding the segmentation step. The proposed framework is validated using the I3A Task-2 dataset over 5-fold cross-validation trials. Using the YOLO predictor, promising mitotic cell prediction results are achieved with an average of 90.011% recall, 88.307% precision, and 81.531% mAP. Whereas, average scores of 86.986% recall, 85.282% precision, and 78.506% mAP are obtained using the Faster R-CNN predictor. Employing the DAL method over four labeling rounds effectively enhances the accuracy of the data annotation, and hence, improves the prediction performance. The proposed framework could be practically applicable to support medical personnel in making rapid and accurate decisions about the mitotic cells' existence.

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

The antinuclear antibody (ANA) test has high sensitivity in diagnosing and classifying systemic lupus erythematosus (SLE).ObjectivesTo 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.MethodsIn 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.ResultsNinety-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.ConclusionThe 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.

Analytical aspects of the antinuclear antibody test by HEp-2 indirect immunofluorescence: EFLM report on an international survey.

Detection of antinuclear antibodies (ANA) by indirect immunofluorescence assay using HEp-2cells (HEp-2 IFA) is used to screen for various autoimmune diseases. HEp-2 IFA suffers from variability, which hampers harmonization. A questionnaire was developed to collect information on HEp-2 IFA methodology, computer-assisted diagnosis (CAD) systems, training, inter-observer variability, quality assessment, reagent lot change control, and method verification. The questionnaire was distributed to laboratories by Sciensano (Belgium), national EASI groups (Italy, Croatia, Portugal, Estonia, Greece) and ICAP (worldwide). Answers were obtained by 414 laboratories. The results were analysed in the framework of the recent EFLM/EASI/ICAP ANA recommendations (companion paper). Laboratories used either HEp-2, HEp-2000, or HEp-20-10cells and most laboratories (80%) applied the same screening dilution for children and adults. The conjugate used varied between laboratories [IgG-specific (in 57% of laboratories) vs. polyvalent]. Sixty-nine percent of CAD users reviewed the automatic nuclear pattern and 53% of CAD users did not fully exploit the fluorescence intensity for quality assurance. Internal quality control was performed by 96% of the laboratories, in 52% of the laboratories only with strongly positive samples. Interobserver variation was controlled by 79% of the laboratories. Limited lot-to-lot evaluation was performed by 68% of the laboratories. Method verification was done by 80% of the respondents. Even though many laboratories embrace high-quality HEp-2 IFA, substantial differences in how HEp-2 IFA is performed and controlled remain. Acting according to the EFLM/EASI/ICAP ANA recommendations can improve the global performance and quality of HEp-2 IFA and nurture harmonization.

The frequency of co-positivity of anti-smooth muscle antibody and anti-nuclear antibodies and their contribution to the diagnosis of autoimmune hepatitis

Aim: Autoimmune hepatitis (AIH) is a chronic disease observed especially in women. The International Autoimmune Hepatitis Group recommends scoring systems for diagnosis using clinical and laboratory data. All scoring systems gave points to autoantibodies as anti-nuclear antibody (ANA) and anti-smooth muscle antibody (SMA) positivity. This study investigates the impact of the co-positivity of the ANA and SMA antibodies on the autoimmune hepatitis diagnosis.&#x0D; Material and Method: We monitored 78 autoimmune liver disease (autoimmune hepatitis, AIH) suspected patients with positive SMA antibody and then further tested for ANA between 2014 and2021. SMA test was screened at 1/40 and 1/100 titers and patients who were positive were taken to further dilution. The ANA test was screened at a titer of 1/40 and 1/160, a positive result was found to be repeated with advanced dilutions. All patients’ autoantibody scores of simplified AIH diagnostic system were calculated.&#x0D; Results: Seventy eight patients with positive SMA antibodies screened for ANA test with 1/40 and 1/160 titer, only 2 patients was found to be negative. The most frequently observed ANA pattern is cytoplasmic linear fibrils (68% ). The 95% ANA positive results was examined at a screening titer of 1/160. The 95% SMA positive results was found at a screening titer of 1/100. The autoantibody scores of 76 patients were +2, patient’s scores were +1.&#x0D; Conclusion: SMA antibody positivity is accompanied by a high rate of ANA antibody positivity but the co-positivity didn’t effect diagnostic score systems. On the other the co-positivity could be a sign of another associated autoimmune diseases.

Differences in the Clinical Significance of Antinuclear Antibodies According to Titers and Patterns in Children.

This study aimed to evaluate the differences in the clinical significance of antinuclear antibody (ANA) according to their titers and patterns in the diagnosis of systemic autoimmune diseases (AiD) in pediatric patients. Of the 2442 children who had undergone an ANA test, 473 (19.4%) were positive for ANA, of whom 33 (7.0%) were diagnosed with significant AiD. The positive predictive value (PPV) for significant AiD was considerably high on application of an ANA titer of ≥1:640, and the PPV of a dense fine speckled (DFS) pattern was significantly lower compared with those of speckled and homogenous patterns. The diagnostic value of ANA positivity for AiD is limited, and the clinical significance of the DFS pattern is relatively lower compared with that of other patterns, such as homogenous and speckled patterns, in children. It is necessary to approach the significance of ANA in children individually depending on titers and patterns.

Evaluation of Antinuclear Antibody and Subserology Reflex Testing for the Diagnosis of Systemic Autoimmune Rheumatic Disorders in an Academic Teaching Hospital.

The aim of this study was to examine appropriate utilization of antinuclear antibody (ANA) screening tests with follow-up subserology tests (reflex testing) for diagnosing systemic autoimmune rheumatic disorder (SARD). We conducted a retrospective chart review of 3003 SARD-test orders at an academic teaching hospital from January to December 2019. Testing patterns were categorized as American College of Rheumatology (ACR)-recommended reflex testing, panel testing, or single subserology testing. We described testing patterns, assessed their diagnostic accuracy, and explored factors associated with reflex testing. Reflex testing accounted for 79.7% of SARD test-ordering, whereas improper testing (panel or single subserology) accounted for the other 20.3%. Reflex testing was associated with significantly more SARD diagnoses than improper testing (P = .004). Testing patterns were significantly associated with race/ethnicity (P = .008), with reflex testing being less frequent than improper testing in Hispanics and Whites. In summary, one-fifth (20.3%) of testing patterns for suspected SARD did not follow the ACR-recommended guidelines for using reflex testing. Use of reflex testing was associated with an increased frequency of SARD diagnosis.

Antinuclear antibodies in children: indirect immunofluorescence patterns, antigen targets and associated diagnoses

Antinuclear antibodies tests are of a paramount importance in the diagnosis, classification, prognostic evaluation and management of autoimmune diseases in children. The present study aimed to describe the immuno-clinical profile of antinuclear antibodies tests in a pediatric population in order to guide the clinical practice of biologists and clinicians. Our study enrolled 268 children. Antinuclear antibodies screening was performed using the indirect immunofluorescence assay on HEp-2 cells. Identification of target antigens was conducted using separately or at one timepoint the following techniques: enzyme-linked immunosorbent assay, Immunodot and Chemiluminescence. The average age of patients was 9.6 ± 4.3 years, with a female predominance (sex-ratio = 1.9). Antinuclear antibodies screening was positive in 40.67% of cases. The most frequently observed antinuclear antibodies patterns were speckled (52.3%), homogeneous (13.8%) and mixed homogeneous-speckled (13.8%). Autoantibodies were detected in 4 patients (2.51%) for whom ANA testing using the indirect immunofluorescence assay was negative. Positive antinuclear antibodies specificities were detected in connective tissue diseases (44.03%; n = 48), organ-specific autoimmune diseases (10.09%; n = 11), and in non-autoimmune conditions (inflammatory diseases, infections, hematological diseases, vasculitis and Wilson’s disease) (32.08%; n = 35). Our study revealed a high rate of positive antinuclear antibodies tests in the pediatric population, mainly related to autoimmune diseases (54.12%) besides non-autoimmune conditions (32.08%). Therefore, screening and interpretation of antinuclear antibodies testing in children require the consideration of clinical data and a close collaboration between clinicians and biologists.

The value of anti-rods and rings antibodies in Chinese population of Dalian City: A retrospective study.

The study aimed to investigate the clinical significance of anti-rods and rings (anti-RR) antibodies in antinuclear antibodies (ANAs) test samples retrospectively. The laboratory data and clinical details of patients with positive anti-RR antibodies were collected and analysed between December 2017 and May 2022 in the First Affiliated Hospital of Dalian Medical University. A total of 72 665 patients were tested for ANAs. There were 45 632 patients discovered with positive ANAs (62.80%), only 131 patients presented with anti-RR antibodies (0.18%), among which only 68 patients were hospitalized patients with a definitive diagnosis. Among the 68 patients with a definitive diagnosis, 8 of 68 (11.8%) had autoimmune diseases, and 19 of 68 (27.9%) had renal diseases. Other diseases included liver disease, pulmonary disease, cerebral ischemia, cerebral infarction, chronic cardiac failure and venous thromboembolism. The detection rate of high titre(≥1:1000) anti-RR antibodies is significantly higher in autoimmune diseases.

Reliability and reproducibility of antinuclear antibody testing in pediatric rheumatology practice.

Antinuclear antibody (ANA) testing is common practice among health care practitioners when evaluating children and adolescents with non-specific symptoms including fatigue and aches and pains. When positive, ANA results often lead to referrals to pediatric rheumatologists as these antibodies may be key indicators for specific pediatric rheumatologic diagnoses. The reliability and reproducibility of ANA tests varies with assay techniques and validation and interpretation of results. In the following article, review of ANA testing in pediatrics is provided along with case examples that demonstrate the reliability and reproducibility of these results in specific scenarios common in the practice of pediatric rheumatology. Guidelines for more accurate utilization of ANA testing are presented with the aim to improve testing and interpretation by ordering clinicians.