Determination of a cut-off value for the serological diagnosis of scrub typhus by detecting anti-Orientia tsutsugamushi immunoglobulin M

Background & objectives: The diagnosis of scrub typhus (ST) is usually done using enzyme-linked immunosorbent assay (ELISA) due to its ease of performance and reading objectivity. The cut-off value for ELISA needs to be calculated for each geographical location as it depends on zonal endemicity of the disease. This study was, therefore, undertaken to calculate the pan-India cut-off for anti-Orientia tsutsugamushi (OT) immunoglobulin M (IgM) by ELISA. Methods: Samples from cases (cases of ST) and controls (voluntary, consenting, healthy adults) were collected by a network of 29 laboratories across India and tested for anti-OT IgM by immunofluorescence assay (IFA), the considered gold standard test. These samples were retested by ELISA for anti-OT IgM and their optical densities (ODs) were used for cut-off estimation by receiver operating characteristic (ROC) curve. Results: Anti-OT IgM ELISA ODs from 273 controls and 136 cases were used for the cut-off estimation. The ODs of the anti-OT IgM ELISA on healthy individuals and those of confirmed ST cases ranged from 0.1 to 0.75 and 0.5 to 4.718, respectively. ROC curve-based cut-off for ELISA was calculated as 0.554 at a sensitivity of 95.2 per cent and specificity of 95.1 per cent. A value of >1 was noted to have a specificity of 100 per cent in diagnosing ST. Interpretation & conclusions: The cut-off calculated for India was similar to the previous cut-off that was used until now.

Utility of oral mucosa as a substrate for the serodiagnosis of pemphigus: A descriptive analysis

Background: The indirect immunofluorescence test is useful in the serodiagnosis of pemphigus. As indirect immunofluorescence titers correlate with disease activity in pemphigus, it is often used as a monitoring tool. The sensitivity of indirect immunofluorescence depends on the substrate used, and the preferred substrates are monkey esophagus for pemphigus vulgaris and normal human skin for pemphigus foliaceus. Aims: We evaluated oral mucosa as a substrate for indirect immunofluorescence in pemphigus. Methods: Fifty patients with pemphigus (40 with pemphigus vulgaris and ten with pemphigus foliaceus) and 50 controls were enrolled for study. Demographic and clinical details were recorded and indirect immunofluorescence using two substrates (oral mucosa and normal human skin) was carried out in serial dilution. Desmoglein (Dsg) 1 and 3 enzyme-linked immunosorbent assay was also evaluated simultaneously. Results: Indirect immunofluorescence was positive in 40 patients (80%) with oral mucosa substrate and 34 patients (68%) with normal human skin substrate. Circulating antibodies were detected with oral mucosa in 33 (82.5%) of the 40 pemphigus vulgaris patients and in 26 (65%) patients using normal human skin. Antibodies were detected in eight of the ten pemphigus foliaceus patients (80%) with normal human skin and in seven (70%) patients with oral mucosa. Dsg enzyme-linked immunosorbent assay was positive in 45 (90%) patients, and 37 of these were also indirect immunofluorescence positive with oral mucosa. In the five Dsg enzyme-linked immunosorbent assay-negative patients, indirect immunofluorescence with oral mucosa was positive in three. Limitations: A comparison of oral mucosa with monkey esophagus could not be performed. Conclusion: Oral mucosa is a suitable and sensitive substrate for indirect immunofluorescence in pemphigus. Further studies comparing the sensitivity of indirect immunofluorescence using oral mucosa with monkey esophagus are recommended.

The influence of demography and referral medical specialty on the detection of autoantibodies to HEP-2 cells in a large sample of patients

Abstract Background: The prevalence of anti-cell autoantibodies detected by indirect immunofluorescence assay on HEp-2 cells (HEp-2-IIFA) increases with age and is higher in female sex. The number of medical specialties that use HEp-2-IIFA in the investigation of autoimmune diseases has increased lately. This study aimed to determine the prevalence and patterns of autoantibodies on HEp-2-IIFA according to demographics variables and referring medical specialties. Methods: A retrospective analysis of the HEp-2-IIFA carried out between January and June of 2017 was performed. The International Consensus on Antinuclear Antibodies Patterns (ICAP) and the Brazilian Consensus on Autoantibodies were used for patterns definition on visual reading of the slides. Anti-cell (AC) codes from ICAP and Brazilian AC codes (BAC) were used for patterns classification. Results: From 54,990 samples referred for HEp-2-IIF testing, 20.9% were positive at titer ≥ 1/80. HEp-2-IIFA positivity in females and males was 24% and 12%, respectively ( p < 0.0001). The proportion of positive results in the 4 age groups analyzed: 0-19, 20-39, 40-59, and ≥ 60 years was 23.3, 20.2, 20.1, and 22.8%, respectively ( p < 0.0001). Considering all positive sera (n = 11,478), AC-4 nuclear fine speckled (37.7%), AC-2 nuclear dense fine speckled (21.3%), BAC-3 nuclear quasi -homogeneous (10%) and mixed/composite patterns (8.8%) were the most prevalent patterns. The specialties that most requested HEp-2-IIFA were general practitioner (20.1%), dermatology (15%), gynecology (9.9%), rheumatology (8.5%), and cardiology (5.8%). HEp-2-IIFA positivity was higher in patients referred by rheumatologists (35.7% vs. 19.6%) ( p < 0.0001). Moderate (46.4%) and high (10.8%) titers were more observed in patients referred by rheumatologists ( p < 0.0001). We observed a high proportion of mixed and cytoplasmic patterns in samples referred by oncologists and a high proportion of BAC-3 (nuclear quasi -homogeneous) pattern in samples referred by pneumologists. Conclusions: One-fifth of the patients studied were HEp-2-IIFA-positive. The age groups with more positive results were 0-19 and ≥ 60 years. AC-4, AC-2, BAC-3 and mixed/composite patterns were the most frequent patterns observed. Rheumatologists requested only 8.5% of HEp-2-IIFA. Positive results and moderate to high titers of autoanti-bodies were more frequent in patients referred by rheumatologists.

Seroprevalence of rickettsial infection in northern Cyprus: A study among hunters

@#This study was conducted to investigate rickettsial seropositivity among hunters, a high-risk population for tick-borne diseases in northern Cyprus. Serum samples were collected from 300 hunters from different locations during the 2017-2018 hunting season (November 2017 - February 2018). The samples were analyzed by indirect immunofluorescence assay (IFA) using slides coated with Rickettsia slovaca, a species belonging to the spotted fever group (SFG). During the sample collection, a questionnaire was also applied to evaluate possible risk factors for rickettsial seropositivity. Of the 300 serum samples, six (2.0%) were found to be IgG-positive with a titer of 1:64. While all seropositive individuals were male, the statistical analysis revealed no significant association of gender with rickettsial seropositivity (p=1.000). Other factors including age (p=0.414), residential places of the participants (p=0.347), hunting years (p=0.694) or hunting abroad (p=1.000) did not significantly affect the IgG positivity. Also, no statistical correlation was found between a history of an arthropod (tick, louse, or flea) bite and rickettsial seropositivity (p=1.000). To our knowledge, this is the first study that demonstrates rickettsial seropositivity among human population in northern Cyprus. Our study suggests that awareness should be raised among the people especially involved in outdoor activities such as hunting, and control programs should be implemented to prevent possible rickettsiosis cases. Further serological studies using other Rickettsia spp. antigens, as well as molecular studies that search for Rickettsia spp. in humans, animals and arthropods are needed to obtain more comprehensive data on rickettsiosis in northern Cyprus.

Evaluation of the value of indirect immunofluorescence on salt-split skin in the diagnosis of bullous pemphigoid / 中华皮肤科杂志

Objective:To evaluate the value of indirect immunofluorescence on salt-split skin (IIF-SSS) in the diagnosis of bullous pemphigoid (BP) .Methods:A single-center clinical retrospective study was conducted. Totally, 163 patients with newly diagnosed BP were collected from Hospital of Dermatology, Chinese Academy of Medical Sciences from January 2013 to January 2019, so were 404 controls, including 161 with pemphigus, 67 with eczema, 26 with drug eruption, 23 with erythema multiforme, 18 with prurigo nodularis, etc. Blood samples were collected before the treatment, and IIF-SSS, BP180 NC16A enzyme-linked immunosorbent assay (ELISA) and direct immunofluorescence (DIF) assay were performed to evaluate the value of IIF-SSS in the diagnosis of BP. Measurement data were compared by using t test and Mann-Whitney test, and enumeration data were compared by using chi-square test and Fisher′s exact test or McNemar test. Results:The number of cases positive for IIF-SSS, BP180 NC16A ELISA and DIF assay was 160, 153 and 127 respectively in the BP group, and 0, 18 and 26 respectively in the control group. The sensitivities of IIF-SSS, BP180 NC16A ELISA and DIF assay for the diagnosis of BP were 98.15%, 93.86% and 77.91% respectively, and their specificities were 100%, 95.54% and 93.56% respectively. There was strong consistency in the diagnosis of BP between IIF-SSS and DIF (Kappa coefficient= 0.767, P < 0.001) . Conclusion:IIF-SSS has relatively high sensitivity and specificity for the diagnosis of BP, and can serve as a routine method for diagnosing BP.

Seroprevalence and hematological abnormalities associated with Ehrlichia canis in dogs referred to a veterinary teaching hospital in central-western Brazil / Soroprevalência e alterações hematológicas associadas com Ehrlichia canis em cães encaminhados a um hospital veterinário-escola no centro-oeste do Brasil

Ticks are significant parasites of dogs in the tropics, where tick-borne pathogens are highly prevalent, especially in areas where tick control measures are frequently neglected. This study investigated the seroprevalence and hematological abnormalities associated with Ehrlichia canis in dogs referred to a veterinary teaching hospital in Central-western Brazil. Out of 264 dogs tested for anti-Ehrlichia canis antibodies by an indirect immunofluorescence assay (IFA), 59.1% (156/264) were positive. Seropositivity was significantly associated to anemia and thrombocytopenia, alone or in combination, and to leukopenia. Conversely, there were no differences in terms of seroprevalence according to sex, breed and age. This study demonstrated that dogs referred to a veterinary teaching hospital in Central-western Brazil are highly exposed to E. canis and that seropositive dogs are more likely to present hematological abnormalities, particularly anemia, thrombocytopenia and leukopenia. To our knowledge, this is the first study on detection of anti-E. canis antibodies by means of IFA among dogs in the state of Goiás. These findings highlighted the need for increasing awareness among dog owners regarding tick control measures in Central-western Brazil, ultimately to reduce the risk of exposure to E. canis and other tick-borne pathogens.

Carrapatos são importantes parasitos de cães nos trópicos, onde patógenos transmitidos por carrapatos são altamente prevalentes, especialmente em áreas onde as medidas de controle de carrapatos são frequentemente negligenciadas. O estudo investigou a soroprevalência e as anormalidades hematológicas associadas à Ehrlichia canis em cães encaminhados para um hospital veterinário-escola no Centro-oeste do Brasil. Dos 264 cães testados para anticorpos anti-Ehrlichia canis por meio da reação de imunofluorescência indireta (RIFI), 59.1% (156/264) foram positivos. A soropositividade foi associada significativamente à anemia e trombocitopenia, isoladamente ou em combinação, e à leucopenia. Por outro lado, não houve diferenças quanto à soroprevalência segundo sexo, raça e idade. Este estudo demonstrou que cães encaminhados a um hospital veterinário-escola na região Centro-oeste do Brasil são altamente expostos à E. canis, e que cães soropositivos têm maior probabilidade de apresentar alterações hematológicas, principalmente anemia, trombocitopenia e leucopenia. Para o nosso conhecimento, este é o primeiro estudo sobre a detecção de anticorpos anti-E. canis por meio da RIFI em cães do estado de Goiás. Essas descobertas destacam a necessidade de aumentar a conscientização entre os proprietários de cães em relação às medidas de controle do carrapato no Centro-oeste do Brasil, em última análise, para reduzir o risco de exposição ao E. canis e outros patógenos transmitidos por carrapatos.

Armonización de la determinación de ANCA por inmunofluorescencia indirecta: primera reunión argentina / Harmonization of the determination of ANCA by indirect immunofluorescence: first argentine meeting / Harmonização da determinação de ANCA por imunofluorescência indireta: primeira reunião argentina

Resumen La determinación de anticuerpos anti-citoplasma de neutrófilos (ANCA) es utilizada en la clínica diaria como una herramienta diagnóstica en distintas formas de vasculitis de pequeños vasos (vasculitis asociadas a ANCA), incluyendo la granulomatosis con poliangeítis (antes: granulomatosis de Wegener), poliangeítis microscópica y granulomatosis eosinofílica con poliangeítis (antes: síndrome de Churg-Strauss), y como apoyo diagnóstico de colitis ulcerosa, colangitis esclerosante primaria y enfermedad de Crohn. Estos anticuerpos están dirigidos contra distintos epítopos antigénicos de diferentes proteínas presentes en el citoplasma del neutrófilo. La determinación de ANCA, habitualmente realizada por la técnica de inmunofluorescencia indirecta (IFI), presenta cierto grado de complejidad en la definición de las imágenes de los diferentes patrones, variabilidad en la forma de trabajo y expresión de resultados. El 24 de octubre de 2018 en el marco del X Congreso Argentino de la Calidad en el Laboratorio Clínico y VIII Jornadas Latinoamericanas de la Calidad en el Laboratorio Clínico, en la Ciudad de Buenos Aires, se realizó una reunión de Armonización de la Determinación de ANCA por IFI con el objeto de presentar, discutir y consensuar los distintos aspectos que se presentan en esta técnica. Las propuestas iniciales fueron discutidas arribándose a recomendaciones generales para proporcionar estándares de trabajo e interpretación de imágenes con el objeto de disminuir la variabilidad de resultados entre los laboratorios clínico-inmunológicos.

Abstract Anti-neutrophil cytoplasmic antibodies (ANCA) tests are widely used in daily clinical practice as a useful tool for the diagnosis of pathologies such as granulomatosis with polyangiitis, microscopic polyangiitis, pauci-immune necrotizing segmental glomerulonephritis and eosinophilic granulomatosis with polyangiitis, ulcerative colitis, primary sclerosing cholangitis, Crohn's disease, etc. These antibodies are directed against different antigenic epitopes of various proteins which are present in the neutrophil cytoplasm. ANCA testing is usually carried out by using indirect immunofluorescence (IIF) method. The determination of ANCA presents some difficulties in the definition of the images of the different patterns, work protocols and result reports uniformity. The "Harmonization Conference on the Determination of ANCA by IIF" was held within the framework of the X Argentine Congress on Quality in the Clinical Laboratory and VIII Latin American Conference on Quality in the Clinical Laboratory, in Buenos Aires, Argentina, on October 24, 2018, in order to present, discuss and agree on different aspects of this method. Some initial proposals were discussed, arriving at general recommendations to provide standards of work and an image interpretation, with the aim of reducing the variability of results among the clinical-immunological laboratories.

Resumo A determinação de anticorpos anti-citoplasma de neutrófilos (ANCA) é utilizada na prática clínica diária como um suporte diagnóstico para várias patologias, como granulomatose com poliangiite, poliangiite microscópica, glomerulonefrite segmentar necrotizante pauci-imune e granulomatose eosinofílica com poliangiite, poliarterite nodosa, colite ulcerosa, colangite esclerosante primária, doença de Crohn. Esses anticorpos são dirigidos contra diferentes epítopos antigênicos de diferentes proteínas presentes no citoplasma do neutrófilo. A determinação de ANCA, normalmente realizada pela técnica de imunofluorescência indireta IFI, apresenta certo grau de complexidade na definição das imagens dos diferentes padrões, variabilidade na forma de trabalho e expressão de resultados. Em 24 de outubro de 2018, no âmbito do X Congreso Argentino de la Calidad en el Laboratorio Clínico (X Congresso Argentino da Qualidade no Laboratório Clínico) e VIII Jornadas Latinoamericanas de la Calidad en el Laboratorio Clínico (VIII Jornadas Latino-americanas da Qualidade no Laboratório), na cidade de Buenos Aires, foi realizada uma Jornada de Harmonização da Determinação de ANCA pelo IFI a fim de apresentar, discutir e concordar sobre os diferentes aspectos que são apresentados nesta técnica. As propostas iniciais foram discutidas, chegando a recomendações gerais para fornecer padrões de trabalho e interpretação de imagens visando reduzir a variabilidade de resultados entre os laboratórios clínico-imunológicos.

Determinación de anticuerpos anti-dsDNA: evaluación del desempeño de un inmunoensayo quimioluminiscente y correlación con el método de inmunofluorescencia indirecta / Detection of anti-dsDNA antibodies: evaluation of performance of a chemiluminescent immunoassay and correlation to the indirect immunofluorescence assay / Determinação de anticorpos anti-dsDNA: avaliação do desempenho de um imunoensaio quimioluminiscente e correlação com o método de imunoflourescência indireta

Resumen La determinación de anticuerpos anti-dsDNA es de utilidad para el diagnóstico y seguimiento clínico de pacientes con lupus eritematoso sistémico (LES) y es uno de los criterios de clasificación del SLICC-2012. El objetivo del estudio fue verificar el desempeño del inmunoensayo quimioluminiscente (CLIA) QUANTA Flash dsDNA y compararlo con el método en uso de inmunofluorescencia indirecta en Crithidia luciliae (CLIFT). Se analizaron con ambos métodos 195 pacientes con diagnóstico de enfermedades del tejido conectivo y solicitud de anticuerpos anti-dsDNA. Se obtuvieron 38 sueros positivos, 133 negativos y 24 (12,3%) discordantes. Entre estos resultados discordantes, hubo 17 que correspondieron a pacientes con LES y se agruparon en 16 CLIA+/CLIFT- y 1 CLIA-/CLIFT+. Se verificó el desempeño para precisión siguiendo el protocolo EP15-A2 y la linealidad. Se estudió la concordancia mediante el coeficiente Kappa y la correlación con el coeficiente Rho de Spearman. Se observó mayor sensibilidad diagnóstica para CLIA. El grado de acuerdo fue moderado y se obtuvo buena correlación entre los valores cuantitativos de CLIA y los títulos de CLIFT. De acuerdo al buen desempeño encontrado y a los resultados discordantes analizados, la mejor estrategia para la implementación de CLIA sería utilizarla en combinación con CLIFT, lo que aumentaría la sensibilidad diagnóstica sin perder especificidad.

Abstract The detection of anti-dsDNA antibodies is useful for diagnosis and clinical monitoring of patients with systemic lupus erythematosus (SLE) and it is part of the classification criteria according to SLICC 2012. The purpose of this study was to verify the performance of the chemiluminescent immunoassay (CLIA) QUANTA Flash dsDNA and to compare it with the method currently in use, i.e the indirect immunofluorescence in Crithidia luciliae (CLIFT). One hundred and ninety five patients, who presented connective tissue diseases and required the study of anti-dsDNA antibodies, were analyzed. Thirty eight positive serum samples were obtained, 133 were negative and 24 (12.3%) in disagreement. Within the discordant results, there were 17 that corresponded to patients with SLE and they were grouped in 16 CLIA+/CLIFT- and 1 CLIA-/CLIFT+. The accuracy performance was assessed according to the EP15-A2 protocol and linearity. Concordance and correlation were calculated with the Kappa and Spearman's Rho coefficient, respectively. Based on the good performance observed and the discordant results analyzed, the best strategy for CLIA implementation would be to combine it with CLIFT, which would increase the diagnostic sensitivity without losing specificity.

Resumo A determinação dos anticorpos anti-dsDNA é de utilidade para o diagnóstico e seguimento clinico de pacientes com lúpus eritematoso sistêmico (LES) e é um dos critérios de classificação do SLICC 2012. O objetivo do estudo foi verificar o desempenho do imunoensaio quimioluminescente (CLIA) QUANTA Flash dsDNA e compará-lo com o método em uso imunofluorescência indireta em Crithidia luciliae (CLIFT). Foram analisados com os dois métodos, 195 pacientes com diagnóstico de doenças do tecido conjuntivo e solicitude de anticorpos anti-dsDNA. Os resultados foram agrupados em 38 soros positivos, 133 negativos e 24 (12,3%) discordantes. Entre esses resultados discordantes, 17 corresponderam a pacientes com LES e se agruparam em 16 CLIA+/CLIFT- e 1 CLIA-/CLIFT+. Foi verificado o desempenho para precisão seguindo o protocolo EP15-A2 e a linearidade. Foi estudada a concordância mediante o coeficiente Kappa e correlação com o coeficiente Rho de Spearman. Observou-se maior sensibilidade diagnóstica para CLIA. O grau de acordo foi moderado e boa correlação foi observada entre os valores quantitativos de CLIA e os títulos de CLIFT. Com base no bom desempenho encontrado e nos resultados discordantes analisados, a melhor estratégia para implementar o CLIA seria utilizá-lo em combinação com o CLIFT, o que aumentaria a sensibilidade do diagnóstico sem perder a especificidade.

Diferencias fenotípicas de fibroblastos gingivales en sujetos con hiperplasia gingival idiopática frente a sujetos periodontalmente sanos: estudio piloto / Phenotypic differences of gingival fibroblasts in subjects with idiopathic gingival hyperplasia versus periodontally healthy subjects: a pilot study

Objetivo: identificar, describir y diferenciar las características fenotípicas de los fibroblastos gingivales (FGs) en pacientes con hiperplasia gingival idiopática (HGI) e individuos periodontalmente sanos. Métodos: los FGs fueron aislados a partir de tejido gingival de individuos periodontalmente sanos (n=2) y pacientes con HGI (n=2). Los FGs se cultivaron en el medio DMEM (Dulbecco's Modified of Eagle Medium) a 37°C con 5% de CO2. La identificación y localización de la actina, vimentina y mitocondrias en FGs fue realizada y evaluada microscópicamente mediante inmunofluorescencia con anticuerpos monoclonales. La capacidad de migración de los FGs en los pacientes con HGI e individuos sanos también fue estudiada. Resultados: todos los FGs fueron mononucleares, fusiformes y con prolongaciones citoplasmáticas visibles. La faloidina permitió identificar una densa red de actina en los FGs de pacientes con HGI, contrariamente a los FGs de individuos periodontalmente sanos. La vimentina y mitocondrias fueron identificadas en los FGs de individuos sanos y pacientes con HGI sin ninguna alteración en su expresión y localización. La migración de la monocapa de los FGs indicó una actividad de migración celular importante en los FGs de los pacientes con HGI, en relación a los FGs de los individuos periodontalmente sanos. Conclusión: los FGs de pacientes con HGI conservan características fenotípicas celulares similares a los FGs de individuos periodontalmente sanos. Sin embargo, los FGs de pacientes con HGI simulan tener una mayor capacidad migratoria que amerita ser explorada en futuros trabajos de investigación.

Objective: To identify and to describe the phenotypic characteristics of gingival fibroblasts from patients with idiopathic gingival hyperplasia (IGH) and periodontally healthy individuals. Methods: Gingival fibroblasts (GFs) were isolated from gingival tissue from periodontally healthy individuals (n=2) and patients with IGH (n=2). The GFs were grown in DMEM (Dulbecco's Modified of Eagle Medium) at 37°C with 5% CO2. The identification and location of actin, vimentin and mitochondria in GFs were performed and evaluated microscopically by immunofluorescence with monoclonal antibodies. The migration capacity of GFs from IGH and healthy individuals was also studied. Results: All the GFs were mononuclear, fusiform and with visible cytoplasmic extensions. The phalloidin allowed to identify a dense actin network in the GFs of patients with IGH, contrary to the GFs of periodontally healthy individuals. Vimentin and mitochondria were identified in the GFs of healthy individuals and patients with IGH without any alteration in their expression and location. Monolayer migration of GFs indicates significant cell migration activity in the GFs of patients with IGH in relation to the GFs of periodontally healthy individuals. Conclusion: GFs from patients with IGH retain cellular phenotypic characteristic similar to GFs from periodontally healthy individuals. However, the GFs of patients with IGH simulate having a greater migratory capacity that deserves to be explored in future research works.

Frecuencia de patrones de anticuerpos antinucleares, en pacientes con sospecha de enfermedades sistémicas reumáticas autoinmunes / Frequency of antinuclear antibodies patterns in patients with suspected of systemic autoimmune rheumatic diseases / Frequência de Padrões de anticorpo antinucleares em pacientes com suspeita de doenças sistêmicas reumáticas autoimunes / Fréquence des profils d'anticorps antinucléaires dans les cas de suspicion de maladies rhumatismales auto-immunes systémiques

Resumen Objetivo: El objetivo de este trabajo es describir la frecuencia de los patrones de tinción de Anticuerpos antinucleares (ANA), en pacientes con sospecha de enfermedades autoinmunes, en el sureste mexicano. Materiales y métodos: Se emplearon células Hep-2 y anticuerpo anti-IgG acoplado a FITC (EuroimmunTM) para el análisis de las muestras a través de inmunofluorescencia indirecta. Resultados: Del total de los pacientes, 89 fueron mujeres (87.2%) y 13 hombres (12.7%), en edades de 2 a 88 años. Se observó que 85 muestras (70.6 %) correspondieron al patrón nuclear, 10 (9.8 %) al patrón citoplasmático y 7 (6.8 %) al patrón mitótico. De los patrones nucleares, 37 (36.8%), 17 (16.7%) y 12 (11.8%) correspondieron a patrones homogéneo, granular fino y granular grueso respectivamente. Conclusiones: En este trabajo se observó, que la mayor frecuencia de anticuerpos antinucleares se encontró en pacientes en edad productiva. Los patrones más observados fueron el homogéneo, granular fino y granular grueso. El patrón homogéneo se asocia a LES cuando se presenta en títulos altos.

Abstract Objective: The objective of this work was to describe the frequency of antinuclear antibody (ANA) staining patterns in patients with suspected autoimmune diseases in southeastern Mexico. Materials and methods: Hep2 cells and Anti-Human IgG coupled to FITC (EuroimmunTM) were used for analyzing samples through indirect immunofluorescence. Results: 87.2 % per cent (89) of the total number of patients were women and 12.7 % were men (13) aged from 2 to 88 years. It was observed that 85 samples (70.6%) corresponded to the nucleolar pattern, 10 (9.8%) to the cytoplasmic pattern and 7 (6.8%) to the mitotic pattern. Of the nuclear patterns, 37 (36.8%), 17 (16.7%) and 12 samples (11.8%) corresponded to homogeneous, nuclear fine speckled and nuclear coarse speckled patterns respectively. Conclusions: In this work, it was observed that the highest frequency of antinuclear antibodies was found in patients of productive age. The most observed patterns were the homogeneous, nuclear fine speckled and nuclear coarse speckled. The homogeneous pattern is associated with SLE when presented in high titers.

Resumo Objetivo: deste trabalho é descrever a frequência dos padrões de coloração de anticorpos antinucleares (ANA) em pacientes com suspeita de doenças autoimunes no sudeste mexicano. Materiais e métodos: células Hep-2 e anticorpo anti-IgG acoplado a FITC (EuroimmunTM) foram utilizados para a análise das amostras por imunofluorescência indireta. Resultados: Do total de pacientes, 89 eram mulheres (87,2%) e 13 homens (12,7%), com idade entre 2 e 88 anos. Observou-se que 85 amostras (70,6%) corresponderam ao padrão nuclear, 10 (9,8%) ao padrão citoplasmático e 7 (6,8%) ao padrão mitótico. Dos padrões nucleares, 37 (36,8%), 17 (16,7%) e 12 (11,8%) corresponderam aos padrões homogêneo, granular fino e granular grosso, respectivamente. Conclusões: Neste trabalho observou-se que a maior frequência de anticorpos antinucleares foi encontrada em pacientes em idade produtiva. Os padrões mais observados foram homogêneo, granular fino e granular grosso. O padrão homogêneo está associado ao LES quando ocorre em altos títulos.

Résumé Objectif : L'objectif de ce travail est de décrire la fréquence des profils de coloration des Anticorps Antinucléaires (ANA), dans les cas de suspicion de maladies auto-immunes, dans le sud-est du Mexique. Matériels et méthodes : Des cellules Hep-2 et un anticorps anti-IgG couplé au FITC (EuroimmunTM) ont été utilisés pour l'analyse desé chantillons par immunofluorescence indirecte. Résultats : Sur le total des patients, 89 étaient des femmes (87,2 %) et 13 des hommes (12,7 %), tous âgés de 2 à 88 ans. Il a été observé que 85 échantillons (70,6 %) correspondaient au patron nucléaire, 10 (9,8 %) au patron cytoplasmique et 7 (6,8 %) au patron mitotique. Parmi es patrons nucléaires, 37 (36,8%), 17 (16,7%) et 12 (11,8%) avaient respectivement un aspect homogène, moucheté à grains fins et moucheté à gros grains. Conclusions : Dans travail, il a été observé que la fréquence la plus élevée d'anticorps antinucléaires a été trouvée chez les patients en âge productif. Les aspects les plus observés ont été de type homogène, moucheté à grains fins et moucheté à gros grains. L'aspect homogène est associé à l'ELS lorsqu'il est présenté en titres élevés.

Detecção de anticorpos anti-Rickettsia rickettsii em cães residentes em área negligenciada no município de São Paulo, SP, Brasil / [Detection of anti-Rickettsia rickettsii antibodies in dogs living in a neglected area in São Paulo, SP, Brazil]

A febre maculosa brasileira (FMB), descrita inicialmente nos Estados Unidos como febre maculosa das Montanhas Rochosas, é uma antropozoonose relatada apenas no continente americano e causada pela bactéria Rickettsia rickettsii. No Brasil a transmissão ocorre sobretudo pela picada de carrapatos do gênero Amblyomma spp. A doença foi inicialmente descrita como de transmissão em áreas rurais e silvestres, no entanto áreas periurbanas e urbanas vêm apresentando casos, principalmente relacionados com a presença de humanos residindo em pequenos fragmentos de mata ciliar. O presente estudo teve por objetivo elucidar a dispersão da FMB nas proximidades dos reservatórios Guarapiranga e Billings, na cidade de São Paulo, SP. Para tanto, a presença de anticorpos anti-R. rickettsii, Rickettsia parkeri e Rickettsia bellii foi avaliada em cães atendidos nas campanhas de esterilização cirúrgica e residentes ao redor dos reservatórios. Foram coletadas amostras de 393 cães, e as amostras de soro foram analisadas pela reação de imunofluorescência indireta (RIFI), com ponto de corte de 1:64. Os títulos para R. rickettsii variaram de 256 a 4096, com positividade de 3,3% (13/393); para R. bellii, de 128 a 1024 e 4,1% (16/393) de positivos, e um único animal (0,25%) foi soropositivo para R. parkeri, com título de 128. Os achados permitem concluir que a região de estudo apresenta condições de se tornar uma possível área com casos de FMB, pois comporta fragmentação de Mata Atlântica, condições essas ideais para a manutenção do vetor do gênero Amblyomma já descrito na região, bem como para a presença da Rickettsia rickettsii circulante entre os cães, confirmada pela existência de anticorpos. Condutas referentes à conscientização da população por meio de trabalhos educacionais devem ser implantadas para a prevenção da doença na população da área.(AU)

Brazilian Spotted Fever (BSF), initially described in the United States as Rocky Mountain Spotted Fever, is an anthropozoonosis reported only in the Americas and caused by the bacterium Rickettsia rickettsii. In Brazil, transmission occurs mainly through tick bites of the genus Amblyomma spp. The disease was initially described as transmission of rural and wild areas; however, peri-urban and urban areas have been presenting cases, mainly related to the presence of humans residing in small fragments of riparian forest. The present study aimed to elucidate the dispersal of BSF near the Guarapiranga and Billings Reservoirs, in the city of São Paulo, SP. The presence of anti-R. rickettsii, Rickettsia parkeri and Rickettsia bellii antibodies were evaluated in dogs treated in surgical sterilization campaigns and residents around the Reservoirs. Samples were collected from 393 dogs and serum samples were analyzed by indirect immunofluorescence reaction (RIFI) with a cutoff of 1:64. The titles for R. rickettsii varied from 256 to 4096 with a positivity of 3.3% (13/393); for R. bellii from 128 to 1024 and 4.1% (16/393) of positive and a single animal (0.25%) was seropositive for R. parkeri with a titre of 128. The findings allow us to conclude that the study region has conditions to become a possible area with BSF cases, as it involves Atlantic Forest, ideal conditions for the maintenance of the vector of the genus Amblyomma already described in the region and the presence of circulating Rickettsia rickettsii among dogs, confirmed by the presence of antibodies. Conducts regarding the awareness of the population through educational work should be implemented to prevent the disease in the population of the area.(AU)

Dinámica de anticuerpos e infección activa por Anaplasma marginale en becerras / Dynamics of Anaplasma marginale Active Infection and Antibodies in Calves

Resumen Históricamente, la especie bovina ha sido de vital importancia para el hombre como animal de abasto, por lo que el cuidado y protección a esta especie es muy importante. A nivel mundial existen diversas enfermedades que atacan el ganado bovino, entre las cuales se encuentra anaplasmosis bovina. La presente investigación no experimental de campo tiene como objetivo evaluar la dinámica de anticuerpos anti-Anaplasma y determinar en qué momento el animal presenta infección activa. Para ello, se escogió una población de 35 becerras ubicadas en la unidad de producción del municipio Crespo, en el estado de Lara, Venezuela. A las becerras, de 0 a 7 días de nacidas, se le extrajo la sangre en tubos con anticoagulante, para realizar frotis de capa blanca y determinación del hematocrito, y sin anticoagulante, para la inmunofluorescencia indirecta. Posteriormente, fueron muestreadas cada 15 días hasta obtener un total de 12 muestras por cada becerra. Para analizar los datos, fue utilizada la prueba estadística X2 del programa estadístico Epi Info versión 3.5.4. Se obtuvo un 79,05 % de positividad y 100 % de seropositividad para Anaplasma marginale. Se concluyó que los altos valores obtenidos con las pruebas serológicas son indicativos de animales con memoria inmunológica, lo que quiere decir que no necesariamente estarían infectados al momento del muestreo. Asimismo, se confirma la capacidad de la especie bovina de crear un estado de premunición ante la anaplasmosis a edades tempranas y mantenerla por un periodo prolongado, de modo que se encuentra una estabilidad enzoótica en la unidad de producción.

Abstract Bovine species has been historically of high importance to the human beings as a provision animal. Therefore, it is highly important to care and protect this species. There are different diseases attacking the bovines around the world, such as the bovine anaplasmosis. This non-experimental research in the field aims to evaluate the dynamics of anti-Anaplasma antibodies and to determine when the animal has an active infection. To do so, a population of 35 female calves was used from the production unit in the town Crespo, Lara State, Venezuela. Female calves, from 0 to 7 days born, were taken blood samples in test tubes with anticoagulant. Then a white smear test was carried out and the hematocrit was determined. Samples without anticoagulant were taken to do an indirect immunofluorescence test. The calves were taken blood samples every 15 days in order to complete 12 samples from each individual calf. For the data analysis, the statistical test X2 from the software Epi Info v.3.5.4 was used. Positive tests accounted for 79.05 % of the samples and seropositiveness for Anaplasma marginale reached 100%. It is concluded that the high values resulting from the serologic tests indicate that these animals have an immune memory, which means, in turn, that they were not necessarily infected at the time of being taken the samples. Likewise, this study confirms that this species is able to create a pre-immunity condition to the anaplasmosis at an early age and to keep it for a long term. This way, they reach an enzootic balance in the production unit.

Rickettsial pathogens circulating in urban districts of Rio de Janeiro, without report of human Brazilian Spotted Fever / Circulação de patógenos riquetsiais em bairros urbanos do Rio de Janeiro sem relatos humanos da Febre Maculosa Brasileira

Abstract Spotted fever group rickettsioses are emerging diseases. In some of these diseases, domestic dogs act as sentinels. Canine serological studies have demonstrated that rickettsial dispersion is concentrated in rural areas, seroprevalence being higher where human rickettsioses are endemic. In Rio de Janeiro, the Atlantic forest vegetation has been devastated by urbanization. In this context, we aimed to detect Rickettsia spp. in urban areas of the West Zone of Rio de Janeiro. Sera from 130 dogs were tested by Indirect Immunofluorescence Assay, and ticks collected from these dogs were tested by polymerase chain reaction. We found the rate of serological reactions against R. rickettsii and R. parkeri in our study area to exceed those of rural and non-endemic areas, highlighting the importance of dogs as urban sentinels. The possibility of contact with opossums and capybaras increased the chances of exposure to Rickettsia spp., reinforcing the hypothetical link between the landscape and the rickettsial wild cycle. Rhipicephalus sanguineus sensu lato was the tick most frequently observed. PCR-positive samples showed similarity with R. rickettsii and R. felis, an emerging pathogen rarely reported from ticks. We observed that rickettsiae circulate in urban places and ticks from indoor environments, which may be involved in bacterial epidemiology.

Resumo Riquetsioses do Grupo da Febre Maculosa são doenças emergentes. Em algumas destas doenças, os cães domésticos agem como sentinelas. Estudos sorológicos caninos têm demonstrado que a dispersão de patógenos rickettsiais está concentrada em áreas rurais, sendo a soroprevalência maior onde as rickettsioses humanas são endêmicas. Na cidade do Rio de Janeiro, a vegetação de Mata Atlântica vem sendo devastada pela urbanização. Nesse contexto, objetivou-se detectar a presença de Rickettsia spp. em áreas urbanas da Zona Oeste do Rio de Janeiro. Amostras de soro obtidas de 130 cães foram testadas, utilizando-se a Imunofluorescência Indireta. Carrapatos coletados desses cães foram testados, utilizando-se a reação em cadeia da polimerase. Observou-se que as taxas de reações sorológicas contra R. rickettsii e R. parkeri nessa área de estudo excederam a prevalência das áreas rurais e não endêmicas, destacando-se a importância dos cães como sentinelas urbanos das rickettsioses. A possibilidade de contato com capivaras e gambás favoreceu a exposição à Rickettsia spp., reforçando a hipótese de ligação entre a paisagem local e o ciclo silvestre de transmissão riquetsial. O carrapato Rhipicephalus sanguineus sensu lato foi encontrado com maior frequência. Amostras com positividade pela PCR mostraram similaridade com R. rickettsii e R. felis, um patógeno emergente raramente descrito em carrapatos. Observou-se circulação riquetsial em áreas urbanas e em carrapatos obtidos do ambiente doméstico, os quais podem estar envolvidos na epidemiologia dessas bactérias.

Molecular diagnosis of a family with May-Hegglin anomaly / 中华医学遗传学杂志

Objective@#To explore the molecular basis for a pedigree affected with May-Hegglin anomaly (MHA).@*Methods@#Peripheral blood samples were collected and subjected to DNA extraction. Exons 1, 10, 16, 24, 25, 26, 30, 31, 33, 38 and 40 and flanking sequences of the MYH9 gene were subjected to PCR amplification and Sanger sequencing. Changes in protein expression were determined by an indirect immunofluorescence assay. Platelet aggregation function of the proband was assessed by thromboelastogram.@*Results@#The proband and his second son both carried a heterozygous 5521G>A (GAG→AAG) missense variant in exon 38 of the MYH9 gene, leading to p. Glu1841Lys substitution at position 1841 of amino acid sequence. Immunofluorescence showed inclusions containing NMMHC-ⅡA. Thromboelastogram suggested enhanced platelet aggregation function of the proband.@*Conclusion@#The c. 5521G>A variant of MYH9 gene has co-segregated with the phenotype of MHA in this pedigree. To assess the aggregation function of platelet by thromboelastogram can predict the risk of bleeding in MHA patients.

Seroprevalence of and Identification of Risk Factors in China / 生物医学与环境科学（英文）

Serum samples were tested for IgG antibodies using indirect immunofluorescence assays. We then analyzed associated risk factors. Serum samples were considered positive when reactive at a dilution of more than 1:320. Differences between groups and risk factors associated with exposure were statistically analyzed using Chi-square tests and the generalized linear model. 122 of 1,260 samples (9.68%) were positive for infection. The infection rate ranged from 0% to 30.43% and differed significantly among age groups ( < 0.01); infection rate in the 50-59 years group was significantly higher than that in other age groups. The seroprevalence of varied significantly among sites within the four provinces, and the infection rate of field workers was significantly higher than that of urban workers.

Low prevalence of infection by Sarcocystis neurona in horses from the State of Alagoas, Brazil / Baixa prevalência da infecção por Sarcocystis neurona em cavalos do estado de Alagoas, Brasil

Abstract The aim of this study was to determine the prevalence of infection by Sarcocystis neurona in horses and identify potential risk factors. Were analyzed 427 samples from 36 farms in 21 municipalities in the Alagoas State, Brazil. Presence of anti-S. neurona antibodies was diagnosed by indirect immunofluorescence antibody test (IFAT) and was confirmed using the immunoblot test. Risk factors were assessed through investigative questionnaires on animal management on the farms. The prevalence of anti-S.neurona antibodies was 2.8% (confidence interval, CI: 1.5-4.9%) from IFAT and 1.6% (CI:0.8-3.34%) from immunoblot, and there were positive horses on 16.6% of the studied farms. None of the variables studied presented associations with serological status for S. neurona. This is the first report on infection by S. neurona in horses reared in Alagoas, Brazil showing a low exposure to S. neurona in this region, but with significant numbers of foci.

Resumo Objetivou-se neste estudo determinar a prevalência e os fatores de risco associados à infecção por Sarcocystis neurona em equinos. Foram analisadas 427 amostras de 36 propriedades localizadas em 21 municípios do estado de Alagoas. O diagnóstico de anticorpos anti-S. neurona foi realizado pela técnica de Imunofluorescência Indireta (IFI) e confirmada por immunoblot. O estudo dos fatores de risco foi realizado a partir de questionários investigativos sobre o manejo dos animais nas propriedades. A prevalência de anticorpos anti-S. neurona foi de 2,8% (I.C. 1,5-4,9%) na IFI e de 1,6% (I.C. 0,8-3,34%) no immunoblot com equinos positivos em 16,6% das propriedades estudadas. Nenhuma variável estudada apresentou associação com o status sorológico para S. neurona. Este é o primeiro relato da infecção por S. neurona em equinos criados no Estado de Alagoas, Brasil, confirmando que os animais desta região têm baixa exposição a S. neurona, mas com significativo número de focos.

Inmunofluorescencia indirecta versus reacción de polimerasa en cadena para el diagnóstico de virus respiratorios en niños ingresados en un hospital de la Región Metropolitana / Indirect immunofluorescence technique versus polymerase chain reaction for the diagnosis of respiratory viruses in children admitted to a hospital in the Metropolitan Region

Resumen Introducción: La temprana detección viral en infecciones respiratorias agudas (IRA) es esencial para establecer una terapia apropiada y prevenir el contagio intrahospitalario. Objetivo: Comparar la eficacia de la técnica de inmunofluorescencia indirecta (IFI) con la reacción de polimerasa en cadena (RPC) para identificar virus respiratorios en niños hospitalizados por IRA. Métodos: Se incluyeron 47 aspirados nasofaríngeos de niños ≤ 2 años con IRA. La IFI incluyó virus respiratorio sincicial (VRS), adenovirus, influenza A y B y parainfluenza. La RPC incluyó, además, la detección de metapneumovirus, enterovirus/rinovirus, bocavirus y coronavirus. Se estimó sensibilidad, especificidad, valor predictor positivo y negativo (VPP/VPN) y correlación kappa para VRS mediante IFI en comparación a la RPC. Resultados: La IFI detectó únicamente VRS (29; 61,7%). La RPC detectó diversos virus, entre ellos VRS en 26 casos (55,3%), seguido por bocavirus (29,8%), enterovirus/ rinovirus (21,3%), adenovirus (14,9%) y parainfluenza (4,3%) entre otros, con 35,5% de co-infección. La IFI presentó sensibilidad: 85,7%, especificidad: 73,6%, VPP: 82,7%, VPN: 77,7% y kappa: 0,5990 (IC 95%; 0,36360,8346) para VRS. Conclusión: La IFI presenta buena sensibilidad, pero moderada especificidad para VRS. Sin embargo, falla en la detección de otros virus respiratorios. La introducción de RPC permitiría mejorar el diagnóstico etiológico de las IRA de origen viral.

Background: Early viral detection in acute respiratory infections (ARI) is essential to establish appropriate therapy and prevent nosocomial transmission. Objective: To compare the efficacy of indirect immunofluorescence technique (IIF) with the polymerase chain reaction (PCR) to identify respiratory viruses in children hospitalized for ARI. Methods: 47 nasopharyngeal aspirates of children ≤ 2 years with ARI were included. IFI included respiratory syncytial virus (RSV), adenovirus, influenza A and B and parainfluenza. PCR also included the detection of metapneumovirus, enterovirus/rhinovirus, bocavirus and coronavirus. Sensitivity, specificity, positive and negative predictive value (VPP/NPV) and kappa correlation for RSV were estimated by IIF compared to PCR. Results: The IIF detected only RSV (29; 61.7%). PCR detected several viruses, including RSV in 26 cases (55.3%), followed by bocavirus (29.8%), rhinovirus/enterovirus (21.3%), adenovirus (14.9%) and parainfluenza (4,3%) among others, with 35.5% of coinfection. The IIF presented sensitivity: 85.7%, specificity: 73.6%, PPV: 82.7%, NPV: 77.7% and kappa: 0.5990 (95% CI, 0.3636-0.8346) for RSV. Conclusion: The IIF presents good sensitivity, but moderate specificity for RSV. However, IIF fails to detect other respiratory viruses. The introduction of PCR would improve the etiological diagnosis of ARI of viral origin.

Concentración sanguínea de troponina I en pacientes embarazadas sin y con preeclampsia severa / Troponin I blood concentration in pregnant patients without and with severe preeclampsia

Resumen OBJETIVO: Comparar la concentración sanguínea de troponina I en pacientes embarazadas sin y con preeclampsia severa. MATERIALES Y MÉTODOS: Estudio prospectivo y transversal efectuado en pacientes de una unidad médica de alta especialidad en Ginecología y Obstetricia de la Ciudad de México admitidas entre los meses de enero a mayo de 2019. Criterio de inclusión: pacientes de cualquier edad y paridad con más de 20 semanas de embarazo. Criterio de exclusión: mujeres con enfermedades previas que pudieran condicionar daño miocárdico. Para cuantificar la troponina I a todas se les tomó sangre venosa. El valor normal de la troponina I está entre 0 y 0.4 ng/mL. Se les tomó un electrocardiograma y una radiografía de tórax. Los reportes de ambos grupos se compararon. Se utilizó estadística descriptiva y prueba t de Student pareada. RESULTADOS: 71 mujeres reunieron los criterios de inclusión y se dividieron en dos grupos: 37 pacientes sin preeclampsia (grupo A) y 34 con preeclampsia (grupo B). La concentración sanguínea de la troponina I del grupo A fue 0.05 ± 0.008 ng/mL y del grupo B: 0.05 ± 0.02 ng/mL (p = 0.30). En el grupo B se encontraron alteraciones electrocardiográficas en 14.7% (n = 5) cuyos valores de troponina I fueron normales. La radiografía del tórax fue normal en 100%. CONCLUSIONES: La concentración sanguínea de troponina I en pacientes embarazadas sin y con preeclampsia severa resultó normal, incluidas las 5 mujeres con preeclampsia y alteraciones electrocardiográficas.

Abstract OBJECTIVE: To compare the blood concentration of Troponin I (TI) in pregnant patients without and with severe preeclampsia (SP). MATERIALS AND METHODS: Prospective, cross-sectional study, conducted in patients of a medical unit of high specialty of gynecology and obstetrics of Mexico City admitted in the period from January to May of the year 2019. Patients of any age and parity with pregnancy > 20 weeks were selected. Those with previous diseases that may condition myocardial damage were excluded. Venous blood was taken to quantify TI using the "Triage Cardiac Panel"® device of the Biosite® brand that runs an indirect immunofluorescence program coupled to a reader called "Triage Meter"®. For this test the normal value of TI is 0 to 0.4 ng/mL. An electrocardiogram and a chest X-ray were also performed. The results of both groups were compared. Descriptive statistics and the paired Student's T test were used. RESULTS: 71 women met the selection criteria. Two groups were formed: 37 patients without preeclampsia (group A) and 34 women with PS (group B). The blood concentration of TI in group A was 0.05 ± 0.008 ng / ml and in group B 0.05 ± 0.02 ng/mL (p = 0.30). In group B electrocardiographic abnormalities were found in 14.70% (5 cases) whose TI values were normal. Chest x-ray was normal in 100%. CONCLUSIONS: the blood concentration of TI in pregnant patients without and with PS was normal, including 5 preeclamptic women with electrocardiographic abnormalities.

V Brazilian consensus guidelines for detection of anti-cell autoantibodies on hep-2 cells

Abstract Background: The V Brazilian Consensus for determination of autoantibodies against cellular constituents on HEp-2 cells, held in Brasilia (DF, Brazil) on August 27, 2016, discussed the harmonization between the Brazilian Consensus on ANA (BCA) guidelines and the International Consensus on ANA Patterns (ICAP) recommendations (www.anapatterns.org). Initial guidelines were formulated by the group of Brazilian experts with the purpose of guiding and enabling Brazilian clinical laboratories to adopt recommendations and to provide a common standard for national and international consensuses. Mainbody: Twenty Brazilian researchers and experts from universities and clinical laboratories representing the various geographical regions of the country participated in the meeting. Three main topics were discussed, namely the harmonization between the BCA guidelines and latest recommendations of the ICAP initiative, the adjustment of the terminology and report on HEp-2 patterns, and a reassessment of quality assurance parameters. For the three topics, our aim was to establish specific guidelines. All recommendations were based on consensus among participants. There was concrete progress in the adjustment of the BCA guidelines to match the ICAP guidelines. To a certain extent, this derives from the fact that ICAP recommendations were largely based on the algorithm and recommendations of the IV Brazilian ANA Consensus, as consistently recognized in the ICAP publications and presentations. However, although there is great overlap between the two Consensuses, there are some point divergences. These specific items were individually and extensively discussed, and it was acknowledged that in several points ICAP improved recommendations previously issued by the Brazilian ANA Consensus and these changes were readily implemented. Regarding some specific topics, the BCA panel of experts felt that the previously issued recommendations remained relevant and possibly will require further discussion with ICAP. The term anti-cell antibodies was adopted as the recommended designation, recognizing that the assay addresses antibodies against antigens in the nucleus and in other cell compartments. However, the acronym ANA HEp-2 was maintained due to historical and regulatory reasons. It was also signalized that the latest trend in ICAP is to adopt the term Indirect Immunofluorescent Assay on HEp-2 cell substrate (HEp-2 IIFA). In addition, the quality assurance strategies previously presented were ratified and emphasized. Conclusion: The V BCA edition was successful in establishing an overall harmonization with the ICAP recommendations for interpretation of the HEp-2 IIFA test, pinpointing the perspectives in filling the remaining gaps between both initiatives.

Circulation of spotted fever group rickettsiae among dogs seropositive for Leishmania spp. in an urban area of Brazil

Abstract INTRODUCTION: Dogs play an epidemiological role in several vector-borne diseases that affect human and animal health worldwide. We aimed to identify rickettsial circulation among dogs with canine visceral leishmaniasis (CVL) from a region endemic for both diseases. METHODS: CVL-seropositive dogs were screened for spotted fever group rickettsiae using an indirect immunofluorescence assay. RESULTS: Among the CVL-positive dogs, anti-Rickettsia rickettsii antibodies were identified in one asymptomatic and one oligosymptomatic dog. CONCLUSIONS: This study shows low circulation of antibodies to R. rickettsii in CVL-seropositive dogs. It is recommended that surveillance studies in dogs should continue in order to monitor this scenario.