scholarly journals Current laboratory and clinical practices in reporting and interpreting anti-nuclear antibody indirect immunofluorescence (ANA IIF) patterns: results of an international survey. 

2020 ◽  
Author(s):  
Lieve Van Hoovels ◽  
Sylvia Broeders ◽  
Edward K.L. Chan ◽  
Luis Andrade ◽  
Wilson de Melo Cruvinel ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Clinical Relevance ◽  
Clinical Practices ◽  
Antibody Testing ◽  
Clinical Value ◽  
International Consensus ◽  
Nuclear Antigens ◽  
Dsdna Antibody ◽  
Homogeneous Pattern ◽  
Rods And Rings

Abstract BackgroundThe International Consensus on Antinuclear Antibody (ANA) Patterns (ICAP) has recently proposed nomenclature in order to harmonize ANA indirect immunofluorescence (IIF) pattern reporting. ICAP distinguishes competent-level from expert-level patterns. A survey was organized to evaluate reporting, familiarity, and considered clinical value of ANA IIF patterns. MethodsTwo surveys were distributed by European Autoimmunity Standardization Initiative (EASI) working groups, the International Consensus on ANA Patterns (ICAP) and UK NEQAS to laboratory professionals and clinicians. Results438 laboratory professionals and 248 clinicians from 67 countries responded. Except for dense fine speckled (DFS), the nuclear competent patterns were reported by >85% of the laboratories. Except for rods and rings, the cytoplasmic competent patterns were reported by >72% of laboratories. Cytoplasmic IIF staining was considered ANA positive by 55% of clinicians and 62% of laboratory professionals, with geographical and expertise-related differences. Quantification of fluorescence intensity was considered clinically relevant for nuclear patterns, but less so for cytoplasmic and mitotic patterns. Combining IIF with specific extractable nuclear antigens (ENA)/dsDNA antibody testing was considered most informative. Of the nuclear competent patterns, the centromere and homogeneous pattern obtained the highest scores for clinical relevance and the DFS pattern the lowest. Of the cytoplasmic patterns, the reticular/mitochondria-like pattern obtained the highest scores for clinical relevance and the polar/Golgi-like and rods and rings patterns the lowest. ConclusionThis survey confirms that the major nuclear and cytoplasmic ANA IIF patterns are considered clinically important. There is no unanimity on classifying DFS, rods and rings and polar/Golgi-like as a competent pattern and on reporting cytoplasmic patterns as ANA IIF positive.

scholarly journals Current laboratory and clinical practices in reporting and interpreting anti-nuclear antibody indirect immunofluorescence (ANA IIF) patterns: results of an international survey

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Lieve Van Hoovels ◽  
Sylvia Broeders ◽  
Edward K. L. Chan ◽  
Luis Andrade ◽  
Wilson de Melo Cruvinel ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Clinical Relevance ◽  
Clinical Practices ◽  
Antibody Testing ◽  
Clinical Value ◽  
International Consensus ◽  
Nuclear Antigens ◽  
Dsdna Antibody ◽  
Homogeneous Pattern ◽  
Rods And Rings

Abstract Background The International Consensus on Antinuclear Antibody (ANA) Patterns (ICAP) has recently proposed nomenclature in order to harmonize ANA indirect immunofluorescence (IIF) pattern reporting. ICAP distinguishes competent-level from expert-level patterns. A survey was organized to evaluate reporting, familiarity, and considered clinical value of ANA IIF patterns. Methods Two surveys were distributed by European Autoimmunity Standardization Initiative (EASI) working groups, the International Consensus on ANA Patterns (ICAP) and UK NEQAS to laboratory professionals and clinicians. Results 438 laboratory professionals and 248 clinicians from 67 countries responded. Except for dense fine speckled (DFS), the nuclear competent patterns were reported by > 85% of the laboratories. Except for rods and rings, the cytoplasmic competent patterns were reported by > 72% of laboratories. Cytoplasmic IIF staining was considered ANA positive by 55% of clinicians and 62% of laboratory professionals, with geographical and expertise-related differences. Quantification of fluorescence intensity was considered clinically relevant for nuclear patterns, but less so for cytoplasmic and mitotic patterns. Combining IIF with specific extractable nuclear antigens (ENA)/dsDNA antibody testing was considered most informative. Of the nuclear competent patterns, the centromere and homogeneous pattern obtained the highest scores for clinical relevance and the DFS pattern the lowest. Of the cytoplasmic patterns, the reticular/mitochondria-like pattern obtained the highest scores for clinical relevance and the polar/Golgi-like and rods and rings patterns the lowest. Conclusion This survey confirms that the major nuclear and cytoplasmic ANA IIF patterns are considered clinically important. There is no unanimity on classifying DFS, rods and rings and polar/Golgi-like as a competent pattern and on reporting cytoplasmic patterns as ANA IIF positive.

scholarly journals Current Laboratory and Clinical Practices in Reporting and Interpreting Anti-nuclear Antibody Indirect Immunofluorescence (ANA IIF) Patterns: Results of an International Survey

2020 ◽  
Author(s):  
Lieve Van Hoovels ◽  
Sylvia Broeders ◽  
Edward K.L. Chan ◽  
Luis Andrade ◽  
Wilson de Melo Cruvinel ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Clinical Relevance ◽  
Clinical Practices ◽  
Antibody Testing ◽  
Clinical Value ◽  
International Consensus ◽  
Nuclear Antigens ◽  
Dsdna Antibody ◽  
Homogeneous Pattern ◽  
Rods And Rings

Abstract BackgroundThe International Consensus on Antinuclear Antibody (ANA) Patterns (ICAP) has recently proposed nomenclature in order to harmonize ANA indirect immunofluorescence (IIF) pattern reporting. ICAP distinguishes competent-levelfrom expert-level patterns. A survey was organized to evaluate reporting, familiarity, and considered clinical value of ANA IIF patterns. MethodsTwo surveys were distributed by European Autoimmunity Standardization Initiative (EASI) working groups, the International Consensus on ANA Patterns (ICAP) and UKNEQASto laboratory specialists and clinicians. Results438 laboratory specialists and 248 clinicians from 67 countries responded. Except for dense fine speckled (DFS), the nuclear competent patterns were reported by >85% of the laboratories. Except for rods and rings, the cytoplasmic competent patterns were reported by >72% of laboratories. Cytoplasmic IIF staining was considered ANA positive by 55% of clinicians and 62% of laboratory specialists, with geographical and expertise-related differences. Quantification of fluorescence intensity was considered clinically relevant for nuclear patterns, but less so for cytoplasmic and mitotic patterns.Combining IIF with specific extractable nuclear antigens (ENA)/dsDNA antibody testing was considered most informative. Of the nuclear competent patterns, the centromere and homogeneous pattern obtained the highest scores for clinical relevance and the DFS pattern the lowest. Of the cytoplasmic patterns, the reticular/mitochondria-like pattern obtained the highest scores for clinical relevance and the polar/Golgi-like and rods and rings patterns the lowest. ConclusionThis survey confirms that the major nuclear and cytoplasmic ANA IIF patterns are considered clinically important. There is no unanimity on classifying DFS, rods and rings and polar/Golgi-like as a competent pattern and on reporting cytoplasmic patterns as ANA IIF positive.

scholarly journals Evaluation of the performance of immunoblot and immunodot techniques used to identify autoantibodies in patients with autoimmune diseases

2021 ◽  
Vol 19 (1) ◽  
pp. 237-244
Author(s):  
Youssef EL Hassouni ◽  
Mohammed Bourhia ◽  
Ahmed Bari ◽  
Riaz Ullah ◽  
Hafiz Majid Mahmood ◽  
...  
Keyword(s):  
Immune System ◽  
Autoimmune Diseases ◽  
Indirect Immunofluorescence ◽  
Antinuclear Antibodies ◽  
Pathological Conditions ◽  
Nuclear Antigens ◽  
Significant Difference ◽  
U1 Snrnp

Abstract Autoimmune diseases are pathological conditions in which the immune system mistakenly attacks its own tissues. This study evaluates the performance of two techniques, which are identifiers of autoantibody specifics: immunoblot and immunodot. This study was conducted in 300 patients of whom 62 were tested positive for antinuclear antibodies. The patients were initially screened for antinuclear antibodies using indirect immunofluorescence. Then, the identification of specific autoantibodies such as anti-extractable nuclear antigens (ENAs) was carried out using the immunoblot and immunodot techniques. The results showed that immunoblot and immunodot did not present a significant difference in their sensitivity against anti-SSA/52, SSB, CENP-B, PCNA, U1-snRNP, Jo-1, Pm-scl, and Mi-2 (p > 0.05). However, the two techniques showed a significant difference in their sensitivity toward autoantibodies anti-DNAn, anti-histone, anti-SmD1, and anti-ds-DNA (p < 0.05). The immunoblot data were in complete accordance with the immunodot data (100%) regarding the detection of autoantibodies such as anti SSA/52, SSB, CENP-B, PCNA, U1-snRP, Jo-1, Pm-scl, and Mi-2, 80% regarding SmD1, and 75% concerning ds-DNA. We should certainly pay closer attention to the efficiency of the techniques used in the diagnosis of autoimmune diseases.

scholarly journals The antinuclear antibody HEp-2 indirect immunofluorescence assay: a survey of laboratory performance, pattern recognition and interpretation

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Anne E. Tebo ◽  
Robert L. Schmidt ◽  
Kamran Kadkhoda ◽  
Lisa K. Peterson ◽  
Edward K. L. Chan ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Clinical Laboratory ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Medical Laboratory ◽  
Nuclear Staining ◽  
International Consensus ◽  
In Vitro Diagnostic ◽  
Traditional Pattern

Abstract Background To evaluate the interpretation and reporting of antinuclear antibodies (ANA) by indirect immunofluorescence assay (IFA) using HEp-2 substrates based on common practice and guidance by the International Consensus on ANA patterns (ICAP). Method Participants included two groups [16 clinical laboratories (CL) and 8 in vitro diagnostic manufacturers (IVD)] recruited via an email sent to the Association of Medical Laboratory Immunologists (AMLI) membership. Twelve (n = 12) pre-qualified specimens were distributed to participants for testing, interpretation and reporting HEp-2 IFA. Results obtained were analyzed for accuracy with the intended and consensus response for three main categorical patterns (nuclear, cytoplasmic and mitotic), common patterns and ICAP report nomenclatures. The distributions of antibody titers of specimens were also compared. Results Laboratories differed in the categorical patterns reported; 8 reporting all patterns, 3 reporting only nuclear patterns and 5 reporting nuclear patterns with various combinations of other patterns. For all participants, accuracy with the intended response for the categorical nuclear pattern was excellent at 99% [95% confidence interval (CI): 97–100%] compared to 78% [95% CI 67–88%] for the cytoplasmic, and 93% [95% CI 86%–100%] for mitotic patterns. The accuracy was 13% greater for the common nomenclature [87%, 95% CI 82–90%] compared to the ICAP nomenclature [74%, 95% CI 68–79%] for all participants. Participants reporting all three main categories demonstrated better performances compared to those reporting 2 or less categorical patterns. The average accuracies varied between participant groups, however, with the lowest and most variable performances for cytoplasmic pattern specimens. The reported titers for all specimens varied, with the least variability for nuclear patterns and most titer variability associated with cytoplasmic patterns. Conclusions Our study demonstrated significant accuracy for all participants in identifying the categorical nuclear staining as well as traditional pattern assignments for nuclear patterns. However, there was less consistency in reporting cytoplasmic and mitotic patterns, with implications for assigning competencies and training for clinical laboratory personnel.

The clinical relevance of anti-dsDNA antibodies determined by the Elia™ dsDNA assay in patients with negative indirect immunofluorescence on the HEp-2 cell

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Christoph Robier ◽  
Maximiliane Haas ◽  
Franz Quehenberger
Keyword(s):  
Indirect Immunofluorescence ◽  
Lupus Erythematosus ◽  
Medical Records ◽  
Clinical Relevance ◽  
Solid Phase ◽  
Clinical Importance ◽  
Discoid Lupus Erythematosus ◽  
Combined Use ◽  
Significant Difference ◽  
Dsdna Antibodies

AbstractObjectivesData on the clinical importance of the detection of anti-dsDNA antibodies in patients with negative indirect immunofluorescence on the HEp-2 cell (IIF) are sparse and are especially not available for all common commercially available assays. This study aimed to assess the clinical significance of anti-dsDNA antibodies determined by the Elia™ dsDNA assay in patients with negative IIF.MethodsWe retrospectively examined the medical records of 234 consecutive subjects with detectable anti-dsDNA antibodies determined by the Elia™ dsDNA assay.ResultsA total of 124 subjects with detectable anti-dsDNA autoantibodies were IIF-negative, but yielded positive or borderline results in the Elia™ CTD screen assay for antinuclear antibodies (ANA). Within this group, 6/49 IIF-negative patients (12%) with ANA-associated systemic autoimmune rheumatic disorders (AASARD) and 118/185 subjects (64%) with various other diseases (Non-AASARD) were identified. There was no statistically significant difference with regard to the concentrations of anti-dsDNA antibodies (p=0.53) between the AASARD and the Non-AASARD group. Within the AASARD group, four patients diagnosed with systemic lupus erythematosus (SLE, treated), discoid lupus erythematosus (untreated), indetermined connective tissue disease (untreated) and polymyositis (treated) had positive anti-dsDNA autoantibodies, whereas two patients with treated SLE, thereby one in remission, had borderline concentrations of anti-dsDNA antibodies.ConclusionsOur findings suggest that the detection of anti-dsDNA antibodies in IIF-negative patients can be of clinical relevance in some cases. Our results further support the combined use of IIF and solid-phase assays in screening algorithms for ANA, in order to avoid overlooking potentially important autoantibody entities.

scholarly journals Assessment of antinuclear antibodies by indirect immunofluorescence assay: report from a survey by the American Association of Medical Laboratory Immunologists

2020 ◽  
Vol 58 (9) ◽  
pp. 1489-1497 ◽  
Author(s):  
Lisa K. Peterson ◽  
Anne E. Tebo ◽  
Mark H. Wener ◽  
Susan S. Copple ◽  
Marvin J. Fritzler
Keyword(s):  
Indirect Immunofluorescence ◽  
Antinuclear Antibodies ◽  
Current Practice ◽  
Clinical Laboratory ◽  
Immunofluorescence Assay ◽  
Indirect Immunofluorescence Assay ◽  
Medical Laboratory ◽  
International Consensus ◽  
Clinical Laboratories ◽  
Reading Interpretation

AbstractBackgroundThe indirect immunofluorescence assay (IFA) using HEp-2 cell substrates is the preferred method by some for detecting antinuclear antibodies (ANA) as it demonstrates a number of characteristic staining patterns that reflect the cellular components bound as well as semi-quantitative results. Lack of harmonized nomenclature for HEp-2 IFA patterns, subjectivity in interpretation and variability in the number of patterns reported by different laboratories pose significant harmonization challenges. The main objectives of this study were to assess current practice in laboratory assessment of HEp-2 IFA, identify gaps and define strategies to improve reading, interpretation and reporting.MethodsWe developed and administered a 24-item survey based on four domains: educational and professional background of participants, current practice of HEp-2 IFA testing and training, gap assessment and the perceived value of International Consensus on Antinuclear Antibody Patterns (ICAP) and other factors in HEp-2 IFA assessment. The Association of Medical Laboratory Immunologists (AMLI) and American Society for Clinical Pathology administered the survey from April 1 to June 30, 2018, to members involved in ANA testing. This report summarizes the survey results and discussion from a dry workshop held during the 2019 AMLI annual meeting.ResultsOne hundred and seventy-nine (n = 179) responses were obtained where a significant number were clinical laboratory scientists (46%), laboratory directors (24%), supervisors (13%) or others (17%). A majority of respondents agreed on the need to standardize nomenclature and reporting of HEp-2 IFA results. About 55% were aware of the ICAP initiative; however, among those aware, a significant majority thought its guidance on HEp-2 IFA nomenclature and reporting is of value to clinical laboratories. To improve ICAP awareness and further enhance HEp-2 IFA assessment, increased collaboration between ICAP and the clinical laboratory community was suggested with emphasis on education and availability of reference materials.ConclusionsBased on these suggestions, future efforts to optimize HEp-2 IFA reading, interpretation and reporting would benefit from more hands-on training of laboratory personnel as well as continuous collaboration between professional organizations, in vitro diagnostic manufacturers and clinical laboratories.

scholarly journals Dose–time fractionation schedules of preoperative radiotherapy and timing to surgery for rectal cancer

2020 ◽  
Vol 12 ◽  
pp. 175883592090753 ◽  
Author(s):  
Fu Jin ◽  
Huanli Luo ◽  
Juan Zhou ◽  
Yongzhong Wu ◽  
Hao Sun ◽  
...  
Keyword(s):  
Rectal Cancer ◽  
Intensity Modulated Radiotherapy ◽  
Clinical Practices ◽  
International Consensus ◽  
Time To Surgery ◽  
Dose Time ◽  
Fractionation Schedule ◽  
Integrated Boost ◽  
Target Volumes ◽  
Fractionation Schedules

Chemoradiotherapy (CRT) is extensively used prior to surgery for rectal cancer to provide significantly better local control, but the radiotherapy (RT), as the other component of CRT, has been subject to less interest than the drug component in recent years. With considerable developments in RT, the use of advanced techniques, such as intensity-modulated radiotherapy (IMRT) in rectal cancer, is garnering more attention nowadays. The radiation dose can be better conformed to the target volumes with possibilities for synchronous integrated boost without increased complications in normal tissue. Hopefully, both local recurrence and toxicities can be further reduced. Although those seem to be of interest, many issues remain unresolved. There is no international consensus regarding the radiation schedule for preoperative RT for rectal cancer. Moreover, an enormous disparity exists regarding the RT delivery. With the advent of IMRT, variations will likely increase. Moreover, time to surgery is also quite variable, as it depends upon the indication for RT/CRT in the clinical practices. In this review, we discuss the options and problems related to both the dose–time fractionation schedule and time to surgery; furthermore, it addresses the research questions that need answering in the future.

scholarly journals Detection of Specific Antinuclear Reactivities in Patients with Negative Anti-nuclear Antibody Immunofluorescence Screening Tests

2002 ◽  
Vol 48 (12) ◽  
pp. 2171-2176 ◽  
Author(s):  
Ilse EA Hoffman ◽  
Isabelle Peene ◽  
Eric M Veys ◽  
Filip De Keyser
Keyword(s):  
Connective Tissue ◽  
Confidence Interval ◽  
Connective Tissue Disease ◽  
Indirect Immunofluorescence ◽  
Connective Tissue Diseases ◽  
Clinical Suspicion ◽  
Screening Tests ◽  
Strongly Correlated ◽  
Nuclear Antigens

Abstract Background: For detection of anti-nuclear antibodies (ANAs) and antibodies to extractable nuclear antigens (ENAs), samples frequently are screened with indirect immunofluorescence (IIF); further determination of anti-ENA antibodies is performed only when the result is positive. However, because anti-ENA reactivities are found in samples with low fluorescence intensities, we determined anti-ENA antibodies in samples with negative IIF and thus calculated the sensitivity of IIF for specific ANAs. Methods: We collected 494 samples consecutively referred by rheumatologists for routine ANA testing. IIF on HEp-2 and HEp-2000 (HEp-2 cells transfected with Ro60 cDNA) and line immunoassay (LIA) for the detection of specific ANAs were performed on all samples. Results: Fluorescence intensities and patterns on HEp-2 were strongly correlated with those on HEp-2000 [Spearman ρ = 0.852 (P &lt;0.001) and 0.838 (P &lt;0.001), respectively]. Sixty-eight of 494 samples were positive on LIA, of which only 72% (confidence interval, 68–76%) were detected with HEp-2 and 75% (confidence interval, 70–78%) with HEp-2000. Of 291 samples negative on both substrates, 12 were positive on LIA. Connective tissue diseases were diagnosed in four of these patients and suspected in at least three others. Conclusion: The HEp-2 and HEp-2000 substrates perform comparably for fluorescence intensities and patterns and for detecting specific ANAs, but some patients with negative IIF show reactivity on LIA. We recommend testing for fine reactivities, regardless of the IIF result, when the clinical suspicion for rheumatic connective tissue disease is high.

scholarly journals Pattern and Frequency of Anti-nuclear Antibody Positivity in Paediatric Rheumatic Diseases

2019 ◽  
Vol 43 (1) ◽  
pp. 21-26
Author(s):  
Mohammad Imnul Islam ◽  
Kamrul Laila ◽  
Shahana A Rahman
Keyword(s):  
Rheumatic Diseases ◽  
Lupus Erythematosus ◽  
Staining Pattern ◽  
Renal Involvement ◽  
Clinical Manifestations ◽  
Speckled Pattern ◽  
Nuclear Antigens ◽  
Antibody Positivity ◽  
Homogeneous Pattern ◽  
Paediatric Rheumatic Diseases

Background: Anti-nuclear antibodies (ANAs) are specific antibodies directed against a variety of nuclear antigens detected in the serum of patients with many rheumatic and non-rheumatic diseases.These antibodies are not only involved in the pathogenesis, but also constitute the basis for diagnosis and treatment of paediatric rheumatic diseases. The objective of the study was to identify the patterns and frequency of ANA positivity in Paediatric Rheumatic Diseases. Methodology: It was a retrospective study. Fourteen hundred and sixty eight records of paediaric rheumatology patients were analyzed. Statistical analysis were done to observe the frequency and association of different patterns of ANA in Juvenile idiopathic arthritis and systemic lupus erythematosus patients. Results: Among the 1468 patients of PRDs, frequency of JIA cases was the highest (65 %) followed by SLE, Scleroderma, juvenile dermatomyositis, and others. Among the 261 PRD patients ANA positivity was 65%. ANA positivity was 100%, 92%, 40% and 31.5% in Mixed connective tissue disease, SLE, JDM and Scleroderma patients respectively. Homogenous staining pattern was found in 59% and speckled pattern in 22.9%. There was significant association between ANA positivity and uveitis in oligoarticular JIA patients. Significant association was also found between homogeneous patterns of ANA and renal involvement in SLE patients. Conclusion: ANA positivity was highest in MCTD cases followed by SLE cases. Majority of SLE cases had homogeneous pattern of ANA.Staining patterns of ANA had significant association with the clinical manifestations in SLE and JIA cases. Bangladesh J Child Health 2019; VOL 43 (1) :21-26

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