Navegando por Palavras-chave "Immunofluorescence"

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    I Consenso Nacional para Padronização dos Laudos de FAN HEp-2
    (Sociedade Brasileira de Patologia ClínicaSociedade Brasileira de PatologiaSociedade Brasileira de Citopatologia, 2002-07-01) Dellavance, Alessandra [UNIFESP]; Gabriel Júnior, Alexandre [UNIFESP]; Cintra, Alice Friedenberg de Ulhôa; Ximenes, Antônio Carlos; Nuccitelli, Barbara; Mühlen, Carlos Alberto Von; Bichara, Carlos David Araújo; Yano, Cristiane; Carvalho, Darlene Gonçalves; Bonfá, Eloisa Silva Dutra de Oliveira; Guimarães, Fabiana N.c.; Mundim, Hugo M.; Pfrimer, Irmtraut Araci Hoffmann; Rego, Jozelia; Andrade, Luiz Eduardo Coelho [UNIFESP]; Mesquita, Mauro Meira de; Santiago, Mittermayer Barreto; Silva, Nilzio Antonio; Miranda, Paulo J.; Leser, Paulo; Francescantonio, Paulo Luiz Carvalho; Jarach, Renata; Levy, Roger Abramino; Neves, Suzane Pretti Figueiredo; Cruvinel, Wilson de Melo [UNIFESP]; Santos, Wilton Silva dos [UNIFESP]; Laboratório Fleury; Centro Imuno Reumatológico de São Paulo; Biorad Diagnósticos; Universidade Federal de Goiás; Laboratório Clínico Padrão; Pontifícia Universidade Católica; Laboratório Amaral Costa; Universidade Católica de Goiás; Laboratório Pardini; Universidade de São Paulo (USP); Laboratório Exame; Universidade Federal de São Paulo (UNIFESP); EBMSP; Universidade Federal de Pernambuco; UERJ; Laboratório Labs; UFMG; Hospital de Base
    The technique of immunofluorescence using HEp-2 cells as substrate is the screening method of choice for the presence of autoantibodies in many clinical laboratories. The lack of a specific terminology for reporting results brings problems in quality control, clinical utility of the test, and standardization attempts. The first Brazilian Consensus for Standardization of ANA in HEp-2 Cells took place in Goiânia in August 2000. Several laboratory specialists with experience in the methodology showed up. They established guidelines for the description of ANA patterns in the Portuguese language, encompassing distinct descriptions for nuclear, nucleolar, cytoplasmic and mitotic apparatus patterns of fluorescence. Recommendations were also established regarding screening titers, final dilution titer, and on morphological criteria for reading the slides.
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    Reprodutibilidade de padrões morfológicos no teste de autoanticorpos contra antígenos celulares (FAN-HEp-2) em diferentes substratos
    (Universidade Federal de São Paulo (UNIFESP), 2020-11-18) Silva, Monica De Jesus [UNIFESP]; Andrade, Luiz Eduardo Coelho [UNIFESP]; Universidade Federal de São Paulo
    The indirect immunofluorescence assay on HEp-2 cells (HEp-2 IFA) is the gold standard method for anti-cellular (AC) antibody screening. Cell culture and fixation methods influence in the distribution and preservation of autoantigens, therefore affect in antigen preservation and distribution, as well as the AC patterns (1,2). Objective: We aimed to evaluate the non-reproducibility phenomenon of HEp-2 IFA results in different HEp-2 slide brands from patients with Systemic Autoimmune Disease (SAD), patients with Non-Autoimmune (NAD), and samples from healthy individuals. Methods: Were evaluated 868 serum samples from 275 patients with SAD, 293 patients with NAD, and 300 samples from healthy blood donors (HBD).Samples were processed at 1:80 dilution according to standard procedure on four HEp-2 slide brands. The tests were interpreted by three experienced independent blinded observers. The agreement among slides was determined using proportional weights according to the number of concordant slides in each sample, yielding a weighted score (ranging from 1 to 100) in specific groups of samples. The qualitative variables were analyzed by the Chi-square test and concordance was analyzed using the Kappa test. All data were analyzed using SPSS20.0 software at a significance level of p <0.05. Results: 402 samples were non-reagent in all slide brands and, therefore, were considered devoid of autoantibodies and eliminated from further analysis. The global reactivity agreement score obtained forth e remaining 466 samples (238 SAD, 119 NAD, and 109 HBD) was considered regular (74, 5). The nuclear compartment showed the highest agreement score (83,6), followed by the metaphase plate (78,9), cytoplasm (77,4) and nucleolus (72,4).Among the clinical groups, the agreement score was higher in SAD (78,0) than in NAD (70,6) and HBD (71,3). Samples with low reactivity (+/4) had higher discordance rates than high intensity samples (++++/4). In the SAD group the discordance for nuclear reactivity was 12.5% for ++++/4 samples, as opposed to 95.6% for +/4 samples, and the same was observed with NAD and HBD samples. The most robust nuclear patterns (higher concordance rate) were, Centromeric (score 78,4), Multiple Nuclear Dots (score73,6), Coarse Speckled (score71,3), and Homogeneous (score 67,9). Among the cytoplasmic patterns, the Reticular Cytoplasmic was more robust (68, 6). Conclusion: The phenomenon of non-reproducibility in the HEp-2 IFA among different slide brands occurs with highest frequency with samples with low intensity.