scholarly journals Properties of Uncommon Indirect Immunofluorescence Staining Patterns Determined during Antinuclear Antibody Detection on HEp-2 Cells

2021 ◽  
Vol 10 (17) ◽  
pp. 3866
Author(s):  
Nada Tomić Sremec ◽  
Ana Kozmar ◽  
Josip Sremec ◽  
Branimir Anić ◽  
Drago Batinić

In this study, we aimed to assess the prevalence of uncommon staining patterns found during testing for the presence of antinuclear antibodies (ANA) and to determine their association with certain antibodies and clinical diagnoses. Presence of ANA and the staining pattern was determined in 10955 samples using indirect immunofluorescence (IIF) on HEp-2 cells. ANA-positive samples were assessed for presence of 14 specific antibody types using a microbead based system. Demographic data (age, sex) and clinical diagnoses were collected from the referral documentation. Particular staining patterns were then compared with a representative comparison group comprised of samples with common staining patterns using these criteria. There were 22 patterns present in less than 3% of samples each and these were jointly present in 42.43% of ANA-positive samples. Specific antibodies were found in proportions similar to the comparison group (46.06%) and varied significantly between patterns. Likewise, there were significant differences in antibody distribution in particular patterns. Some patterns were associated with presence of rheumatic diseases or inflammatory arthropathies, while in others there was a concurrent diagnosis of liver disease, or a neoplastic process. Many of the uncommon IIF patterns have distinctive characteristics that warrant further investigation in order to determine their role in diagnosing various diseases, not limited only to the illnesses of the rheumatic spectrum. IIF on HEp-2 cells remains an irreplaceable method because of the diversity of ANA, only a number of which can be detected using other standardised methods.

Author(s):  
Xavier Bossuyt ◽  
Lisette Meurs ◽  
Alex Mewis ◽  
Godelieve Mariën ◽  
Norbert Blanckaert

We evaluated indirect immunofluorescence (IF) using HEp-2000TM slides, which are transfected with SS-A cDNA, for screening for anti-SS-A antibodies, by comparing it with counterimmunoelectrophoresis (CIE). A total of 2427 specimens were screened for IF reactivity and for SS-A precipitins, of which 1033 (43%) were negative on both IF and CIE. There were 1271 SS-A precipitin-negative specimens (52%) which were IF-positive but lacked the distinctive SS-A staining pattern. One precipitin-negative serum was IF-positive with the distinctive SS-A pattern in the HEp-2000 system. One hundred and twenty-two specimens (5%) were positive for anti-SS-A precipitins on CIE, 107 showed the distinctive SS-A fluorescence staining pattern, whereas 15 of these precipitin-positive samples (12%) were IF-positive but did not display the distinctive SS-A pattern on the transfected cells. Fourteen of the 15 samples in which the distinctive SS-A pattern was not observed displayed other significant antinuclear antibody (titre equal or > 1:320 patterns. In conclusion, the presence of the typical 'distinctive' SS-A pattern on IF using the HEp-2000 slides is highly specific for the presence of autoantibodies to SS-A and has a sensitivity of 88% for detecting these antibodies.


2018 ◽  
Vol 78 (6) ◽  
pp. e48-e48 ◽  
Author(s):  
Lieve Van Hoovels ◽  
Sofie Schouwers ◽  
Stefanie Van den Bremt ◽  
Xavier Bossuyt

2021 ◽  
Author(s):  
Zeki Yumuk ◽  
Elif Okumuş

Abstract We examined correlates of antinuclear antibody (ANA) positivity in individuals with COVID-19 infection. Sera from 156 patients were evaluated by indirect immunofluorescence on Hep-2 cells for ANA pattern and semi-quantitative titer. There was a significant difference in the prevalence of ANA (1:100) by gender (P = 0.0294). Male gender was correlated with ANA positivity and association persisted in multivariable analyses (OR = 0.354; CI: 0.139–0.816; P = 0.0200). Age (OR = 1.932; CI: 0.929–4.121; P = 0.0814), warm weather (OR = 1.307; CI: 0.611–2.921; p = 0.4989), geographic location (or = 0.886; CI:0.353–2.442; P = 0.8042), and diabetes mellitus (OR = 1.765; CI:0.513–5.477; P = 0.3369) were not correlated with ANA positivity. In this analysis, hospitalization was used as indicator for a more severe course. Of the 156 COVID-19 patients, 18 (11.5%) were hospitalized. Among 18 hospitalized patients, 4 (22.2%) were ANA positive (Table 2), although this did not reach statistical significance (OR = 0.841; CI: 0.227–2.527; p = 0.7725). In conclusion, although males were more prone to produce ANA during COVID-19 infection, this was not correlated with severe course.


2021 ◽  
Vol 19 (1) ◽  
pp. 237-244
Author(s):  
Youssef EL Hassouni ◽  
Mohammed Bourhia ◽  
Ahmed Bari ◽  
Riaz Ullah ◽  
Hafiz Majid Mahmood ◽  
...  

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.


2020 ◽  
Vol 58 (9) ◽  
pp. 1489-1497 ◽  
Author(s):  
Lisa K. Peterson ◽  
Anne E. Tebo ◽  
Mark H. Wener ◽  
Susan S. Copple ◽  
Marvin J. Fritzler

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.


2011 ◽  
Vol 10 (12) ◽  
pp. 801-808 ◽  
Author(s):  
Katrijn Op De Beeck ◽  
Pieter Vermeersch ◽  
Patrick Verschueren ◽  
René Westhovens ◽  
Godelieve Mariën ◽  
...  

1998 ◽  
Vol 15 (2) ◽  
pp. 7-38 ◽  
Author(s):  
Ann Sanson ◽  
Margot Prior ◽  
Frank Oberklaid ◽  
Diana Smart

AbstractResults are presented from a recent study within the Australian Temperament Project (ATP), in which a group of children with significant behaviour problems, and a comparison group, were selected from the sample at 11–12 years and home-visited, with assessments of clinical diagnoses, intelligence, school achievement and social competence, and a variety of family functioning indices. Approximately half the behaviour problem group received at least one diagnosis. Twice as many boys as girls were diagnosed. Rates of comorbidity were high but, generally, within—rather than between—the broadband internalising or externalising spectra. Concurrent family functioning measures discriminated between groups, but not as strongly as intrinsic child measures, and the particular family variables that best discriminated between groups showed sex differences. High stability of behaviour problems from earlier years was evident, and the behaviour problem group differed from the comparison group on measures of temperament, behaviour, and context from early childhood; both findings reinforce the need for early intervention.The implications of these and other findings from the ATP, particularly the need for early intervention, are discussed.


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