ABSTRACT
Resumen Objetivo: Determinar la frecuencia de anticuerpos antimitocondriales y de anticuerpos contra antígenos extraíbles del núcleo en pacientes con cirrosis biliar primaria. Material y métodos: Estudio de tipo cuantitativo, observacional y transversal, realizado en el Servicio de Inmunología del Hospital Nacional Arzobispo Loayza entre enero 2018 y marzo 2019. Se revisaron las historias clínicas de 30 pacientes con características presuntivas de cirrosis biliar primaria; para la detección de los anticuerpos antinucleares y anticuerpos antimitocondriales se empleó el kit inmunológico en sangre y observación con microscopio de inmunofluorescencia a 40X y para la detección de los anticuerpos contra antígenos extraíbles del núcleo se empleó el método Immunoblot. Resultados: Se estudiaron 30 pacientes con cirrosis biliar primaria, 20 fueron de sexo femenino (66,7%). El patrón de tinción más frecuente fue el citoplasmático moteado reticular en 17(56,7%), seguido del patrón citoplasmático moteado reticular y patrón moteado en 7(23,3%) pacientes, y en menor frecuencia el patrón citoplasmático moteado reticular y patrón centromérico. Nueve (42,9%) pacientes con cirrosis biliar primaria tenían anti-M2. Se demostró mayor frecuencia, 21(70%) de los pacientes con cirrosis biliar primaria tenían anticuerpos antimitocondriales. Conclusiones: Se encontró alta frecuencia de patrón citoplasmático moteado reticular en pacientes con cirrosis biliar primaria, se demostró asociación significativa con los anti-M2 y anticuerpos antimitocondriales.
Summary Objective: To determine the frequency of antimitochondrial antibodies and antibodies against extractable nucleus antigens in patients with primary biliary cirrhosis. Methods : A quantitative, observational and cross-sectional study was carried out at the Immunology Service of the Arzobispo Loayza National Hospital between January 2018 and March 2019. The medical records of 30 patients with presumptive characteristics of primary biliary cirrhosis were reviewed; for the detection of the antinuclear antibodies and antimitochondrial antibodies, the immunological kit was used in blood and observation with a 40X immunofluorescence microscope, and the Immunoblot method was used for the detection of the antibodies against extractable nucleus antigens. Results: Thirty patients with primary biliary cirrhosis disease were studied, 20 were female (66.7%). The most frequent staining pattern was the reticular mottled cytoplasmic in 17 (56.7%), followed by the reticular mottled cytoplasmic pattern and mottled pattern in 7 (23.3%) patients, and less frequently the reticular mottled cytoplasmic pattern and centromeric. Nine (42.9%) patients with primary biliary cirrhosis had anti-M2. In the present investigation, a higher frequency was demonstrated, 21 (70%) of the patients with primary biliary cirrhosis had antimitochondrial antibodies. Conclusions: A high frequency of reticular mottled cytoplasmic pattern was found in patients with primary biliary cirrhosis; a significant association with anti-M2 and antimitochondrial antibodies was demonstrated.
ABSTRACT
ABSTRACT Introduction: The detection of anti-double-stranded (ds) deoxyribonucleic acid (DNA) antibodies is one of the classification criteria for diagnosing systemic lupus erythematosus (SLE). Objective: To describe a quantitative enzyme-linked immunosorbent assay (ELISA) for detecting anti-dsDNA immunoglobulin class G (IgG) antibodies. Methods: The performance of ELISA was evaluated using the Crithidia luciliae indirect immunofluorescence test (CLIFT) as a reference. Anti-dsDNA IgG antibodies were screened by ELISA and CLIFT in serum samples from 127 patients with SLE, 56 patients with other diseases and 37 healthy persons. The Cochran Q test was used to compare the sensitivity and specificity of the reactions, with differences among the results being considered significant when p ≤ 0.05. Results: ELISA had a sensitivity of 92.9% and a specificity of 94.6%, whereas the sensitivity and specificity of CLIFT were 85.8% and 100%, respectively. ELISA was significantly more sensitive than CLIFT (p = 0.0027), whereas CLIFT was significantly more specific than ELISA (p = 0.0253). Conclusion: ELISA showed excellent results in terms of sensitivity and specificity, with a potential use in research and routine diagnostics.
RESUMEN Introducción: La detección de anticuerpos contra el ácido desoxirribonucleico (ADN) de doble cadena (dc) es uno de los criterios de clasificación para el diagnóstico de lupus eritematoso sistémico (LES). Objetivo: Describir una técnica inmunoenzimática (ELISA) cuantitativa para detección de anticuerpos de inmunoglobulina de clase G (IgG) anti-ADNdc. Métodos: Se evaluó el desempeño de la técnica ELISA mediante el test inmunofluorescencia indirecta con Crithidia luciliae (IFI-CL) como referencia. Anticuerpos IgG anti-ADNdc fueron analizados por ELISA y IFI-CL en muestras de sueros de 127 pacientes con LES, 56 pacientes con otras enfermedades y 37 personas sanas. La prueba Q de Cochran fue utilizada para comparar la sensibilidad y la especificidad de las reacciones considerando diferencias significantes entre los tests cuando p ≤ 0,05. Resultados: La técnica ELISA mostró sensibilidad del 92,9% y especificidad del 94,6%, mientras la sensibilidad y la especificidad de la técnica IFI-CL fueron del 85,8% y 100%, respectivamente. La técnica ELISA mostró sensibilidad significativamente mayor que la obtenida con IFI-CL (p = 0,0027); esta mostró especificidad significativamente mayor que la obtenida con ELISA (p = 0,0253). Conclusión: La técnica ELISA presentó resultados excelentes de sensibilidad y especificidad, con el potencial de ser utilizada en investigación y rutina diagnóstica.
RESUMO Introdução: A detecção de anticorpos contra o ácido desoxirribonucleico (DNA) nativo (ds) é um dos critérios de classificação para o diagnóstico do lúpus eritematoso sistêmico (LES). Objetivo: Descrever uma técnica imunoenzimática enzyme-linked immunosorbent assay (ELISA) quantitativa para a detecção de anticorpos imunoglobulina da classe G (IgG) anti-DNAds. Métodos: O desempenho da técnica ELISA foi avaliado utilizando o teste de imunofluorescência indireta com Crithidia luciliae (CLIFT) como referência. Anticorpos IgG anti-DNAds foram pesquisados por ELISA e CLIFT em amostras de soros de 127 pacientes com LES, 56 pacientes com outras doenças e 37 indivíduos sadios. O teste Q de Cochran foi utilizado para comparar as sensibilidades e as especificidades das reações, considerando diferenças significantes entre os testes quando p ≤ 0,05. Resultados: A técnica ELISA apresentou sensibilidade de 92,9% e especificidade de 94,6%, enquanto a sensibilidade e a especificidade da técnica CLIFT foram de 85,8% e 100%, respectivamente. A técnica ELISA apresentou sensibilidade significativamente maior do que a obtida com a técnica CLIFT (p = 0,0027); esta apresentou especificidade significativamente maior do que a obtida com a técnica ELISA (p = 0,0253). Conclusão: A técnica ELISA apresentou excelentes resultados em termos de sensibilidade e especificidade, podendo ser útil em pesquisa e rotina diagnóstica.
ABSTRACT
ABSTRACT Introduction: The detection of autoantibodies in HEp-2 cells represents a relevant tool for the diagnosis of autoimmune diseases, especially rheumatic autoimmune diseases. As a result of the methodological advances, the technique gradually increased the sensitivity, as well as the need for standardization. Objective: To evaluate the implementation of the Brazilian Consensus recommendations for autoantibody determination in HEp-2 cells. Methods: A structured form in a virtual platform was filled in by experts in clinical laboratories that carry out the methodology across the country. The questionnaire addressed the adoption of the Brazilian Consensus guidelines, detailing the technical aspects, quality control, the strategy for reading slides and the release of reports. Results: The study included 53 laboratories responsible for more than 300,000 antinuclear antibody (ANA) tests/month; more than half (58.5%) reported fully adopting the recommendations of the Brazilian Consensus. The majority (83.1%) used the 1:80 for screening dilution, and 75.5% of laboratories, perform education and quality control programs. Only 39.6% reported using more than one kit brand to perform the test, and 32.1% did not report observing all phases of the cell cycle during slide reading. The study also detected some heterogeneity among participants in the identification of patterns. Conclusion: The results confirm the adoption of the Brazilian Consensus recommendations by most of participating laboratories, although with variable extent. There is need for improvement in some aspects, especially those related to the quality control.
RESUMO Introdução: A pesquisa de autoanticorpos em células HEp-2 representa uma relevante ferramenta no auxílio diagnóstico de doenças autoimunes, especialmente as reumáticas. Em virtude dos avanços metodológicos, a técnica aumentou gradativamente a sensibilidade, bem como a necessidade de padronização. Objetivo: Avaliar a implantação das recomendações dos consensos brasileiros de pesquisa de autoanticorpos em células HEp-2. Métodos: Preenchimento de formulário em plataforma virtual direcionada aos laboratórios clínicos que realizam a metodologia. Os participantes responderam a um questionário sobre a adoção das diretrizes dos consensos brasileiros, detalhando os aspectos técnicos, o controle de qualidade, a leitura de lâminas e a emissão de laudos. Resultados: Participaram do estudo 53 laboratórios responsáveis por mais de 300 mil testes de fator antinuclear (FAN)/mês; mais da metade (58,5%) informou adotar integralmente as recomendações dos consensos. A maioria (83,1%) utiliza a diluição 1:80 para triagem, e 75,5% dos laboratórios, programas de educação e controle de qualidade. Apenas 39,6% utilizam mais de uma marca de kit para a realização do teste, e 32,1% não relataram observar todas as fases do ciclo celular na leitura da lâmina. O estudo detectou ainda discreta heterogeneidade entre participantes na identificação de padrões. Conclusão: Os resultados evidenciam a adoção das recomendações dos consensos de forma absoluta pela maioria dos laboratórios participantes, bem como a necessidade de aperfeiçoamento em alguns aspectos relevantes para a qualidade do ensaio.
ABSTRACT
A 66-year-old man with a history of multiple transurethral resections for recurrent bladder tumors, staged as Ta according to the International Union Against Cancer staging guidelines, presented with a complaint of dry cough. A round nodule with a diameter of 7.5 cm was detected in the lung by chest computed tomography, and a video-assisted thoracoscopic lobectomy was performed. Pulmonary metastasis of recurrent bladder cancer was diagnosed by immunohistochemistry staining for the urothelium-specific protein uroplakin Ia. Subsequently, 2 cycles of systemic chemotherapy were administered. Two and a half years after treatment, no recurrence of pulmonary lesions has been detected. A combination of complete resection of pulmonary lesions and systemic chemotherapy may result in a good prognosis for patients with non-muscle-invasive bladder cancer.
Subject(s)
Humans , Cough , Fluorescent Antibody Technique, Indirect , Immunohistochemistry , Lung , Neoplasm Metastasis , Neoplasm Staging , Prognosis , Recurrence , Solitary Pulmonary Nodule , Thorax , Urinary Bladder , Urinary Bladder Neoplasms , Uroplakin IaABSTRACT
El objetivo de este estudio fue determinar los niveles de anticuerpos posvacunales contra diferentes serovares de Leptospira en 78 cerdas de cría en dos momentos diferentes de la gestación (días 90 a 95: momento 1 y días 110 a 114: momento 2), mediante el uso de las técnicas de Microaglutinación (MAT) e Inmunofluorescencia Indirecta (IFI). Cada suero fue probado contra seis serovares de Leptospira (Canicola, Hardjo bovis, Pomona, Grippotyphosa, Icterohaemorrhagiae y Bratislava). Los resultados obtenidos muestran un 59% de positividad mediante MAT en el primer muestreo y del 17% en el segundo muestreo, siendo los serovares Icterohaemorrhagiae y Hardjo bovis los más prevalentes; por medio de IFI se encontró un 100% de sueros positivos en ambos muestreos para al menos uno de los serovares estudiados, con títulos entre 1:64 y 1:512, en el primer muestreo y una pronunciada disminución en los títulos en el segundo muestreo o negativización de los mismos; los serovares más prevalentes por IFI fueron Icterohaemorrhagiae y Canicola. Se concluye de esta investigación la baja capacidad protectora de esta vacuna al encontrar un aumento en el número de animales seropositivos para IgM en el segundo muestreo frente a algunos de los serovares estudiados, así como también el aumento de los títulos de IgG frente a Icterohaemorrhagiae en el segundo muestreo y su pobre capacidad de protección a los lechones en sus primeros días de vida al observarse una negativización de ambos tipos de anticuerpos al final de la gestación.
The goal of this study was to determine the levels of postvaccinal antibodies against several serovars of Leptospira in 78 sows in two different gestation moments (day 90 to 95: moment 1, and day 110 to 114: moment 2), using the Micro agglutination (MAT) and Indirect Immunofluorescence (IFI) techniques. Each serum was tested for six serovars of Leptospira (Canicola, Hardjo bovis, Pomona, Grippotyphosa, Icterohaemorrhagiae and Bratislava) donated by the Center of Diagnostics of the Instituto Colombiano Agropecuario (ICA) in Manizales-Caldas-Colombia. The results showed a 59% positive sera using MAT in moment one and a 17% in moment two, with Icterohaemorrhagiae and Hardjo bovis as the most prevalent. By means of the IFI technique 100% of sera were positive in both moments, for at least one of the serovars tested, with titers between 1:64 and 1:512 in the first moment and a strong decrease or absence of the titers in the second moment; the most prevalent serovars for IFI were Icterohaemorrhagiae and Canicola. In conclusion, this vaccine has a low protection capacity since an increase in the number of seropositive sows with IgM was observed against some serovars of Leptospira in the second moment, as well as an increase in the levels of IgG against Icterohaemorrhagiae in the second moment, and its low protection capacity in offspring in their first days of life associated with a decrease of the antibodies at the end of the gestation.