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.

Brote urbano de leishmaniasis visceral en Neiva, Colombia / Urban outbreak of visceral leishmaniasis in Neiva (Colombia)

Objetivo Estudiar clínica y epidemiológicamente focos de leishmaniasis visceral (LV) urbana en Neiva (Colombia). Materiales y Métodos Seis niños consultaron por hepato-esplenomegalia. Presentaban anemia y leucopenia. Se realizó biopsia por aspiración de medula ósea (5 pacientes) y de bazo (1 paciente). Se hizo búsqueda activa de casos en la comunidad y de anticuerpos anti-Leishmania infantum por inmunofluorescencia indirecta (IFI) en los sintomáticos y en reservorios caninos (IFI, rK39). Se hicieron visitas domiciliarias para educación comunitaria y búsqueda de vectores. Los pacientes recibieron Miltefosina, Anfotericina B o Glucantime®. Resultados Se confirmó LV en siete niños. En seis, el aspirado de medula ósea o bazo demostró amastigotes. La IFI fue positiva en 4 pacientes y negativa en 3. Un niño se detectó por búsqueda activa comunitaria, con clínica de LV, confirmada por IFI (1:32). La miltefosina no fue útil en 6 de los 7 casos. La Anfotericina B liposomal o deoxicolato, curó 6 pacientes y el Glucantime® uno. La seroprevalencia en 1182 caninos (IFI y rK39) fue de 6.1 %; los animales positivos fueron sacrificados. Se demostró Lu. longipalpis, vector de LV, en el peridomicilio. Conclusiones Demostramos LV urbana en dos comunas de Neiva. La confirmación diagnóstica incluyó aspiración de medula ósea e IFI. La Miltefosina no fue útil. La Anfotericina B liposomal fue la terapia ideal. Para controlar la LV es necesario hacer BAC, educación comunitaria, control de vectores y reservorios.(AU)

Objective Characterize the foci of visceral leishmaniasis infection in Neiva with a clinical and epidemiological approach. Materials and Methods Six children consulted medical services with hepatosplenomegaly. They were found to have anemia and leukopenia. The diagnosis was performed by bone marrow (five patients) and spleen (1 patient) aspiration. An active search for cases was carried out in the community. Anti-Leishmania infantum antibodies were also sought out using indirect immunofluorescence (IIF) in symptomatic patients and in dogs (IFI, rK39). House calls were made in order to carry out educational activities and to collect disease vectors. Patients received miltefosine, amphotericin B or Glucantime®. Results LV was confirmed in seven children. In six of them, the bone marrow or spleen aspirate contained amastigotes. The IIF was positive in 4 patients and negative in 3. One child was detected throught the active community search, confirmed by the clinic with IIF (1:32). Six patients were cured with liposomal amphotericin B (o deoxycholate) and one patient was cured with Glucantime®. The canine seroprevalence in 1182 dogs was 6.1% (IFI and rK39); the positive animals were destroyed. L. longipalpis was found in the houses. This is the principal vector of LV in Colombia. Conclusions The study showed that two zones of Neiva have children infected with LV. Diagnostic confirmation must include aspiration of bone marrow and IIFs. Treatment with miltefosine was not helpful, but liposomal amphotericin B is an ideal therapy. To control LV, active case searching, community education and vector and reservoir control is necessary.(AU)

Diagnóstico da Leishmaniose visceral canina: avalição do desempenho dos kits EIE-Leishmaniose-visceral-canina-Bio-Manguinhos e IFI-Leishmaniose-visceral-canina-Bio-Manguinhos / Diagnosis canine visceral Leishmaniasis: perfomance evaluation of EIE-Leishmaniose-visceral-canina-Bio-Manguinhos and IFI-Leishmaniose-visceral-canina-Bio-Manguinhos kits

Avaliou-se o desempenho do kit EIE-Leishmaniose-Visceral-Canina-Bio-Manguinhos (EIE-LVC) e foi comparado seu desempenho com o kit IFI-Leishmaniose-Canina-Bio-Manguinhos (IFI-LVC), visando sua utilização em rotina epidemiológica. Foram utilizados 114 cães atendidos no Hospital Veterinário da UFRPE. Constitui-se quatro grupos: Grupo 1 (G1), animais com sinais clínicos sugestivos de LVC e com testes parasitológicos positivos (n=25); Grupo 2 (G2), animais com diagnóstico apenas presuntivo de LVC (n=62); Grupo 3 (G3), animais que nunca habitaram em área endêmica para a LVC, nunca receberam transfusão sanguínea (n=16); e Grupo 4 (G4), animais portadores de outros etiologias: babesiose (n=4), anaplasmose (n=6) e demodicose (n=1). A análise do EIE-LVC no G1 apresentou uma sensibilidade de 72,0 por cento e especificidade de 87,0 por cento. Os valores de VPP e VPN foram de 90,0 e 66,7 por cento, respectivamente. O valor do índice kappa foi de 0,975. Quando esses grupos foram analisados através do IFI-LVC, no G1, a sensibilidade foi de 68,0 por cento e a especificidade de 87,5 por cento. Os valores de VPP e VPN foram de 89,5 e 63,6 por cento, respectivamente. (...) foi evidenciado que os testes sorológicos não se mostraram estatisticamente diferentes quanto a performance para detecção de anticorpos anti-Leishmania sp. Quando as amostras foram analisadas em paralelo a sensibilidade foi de 92,0 por cento e a especificidade de 75,0 por cento. Já na análise dos testes em série a sensibilidade foi de 48,0 por cento e a especificidade de 100,0 por cento. A análise do G2 através dos kits de IFI-LVC e EIE-LVC em paralelo, revelou que neste grupo 26 animais foram detectados como positivos e 36 foram diagnosticados como negativos. A análise do diagnóstico clínico frente à associação dos testes em paralelo apresentou um VPP de 42,0 por cento. Quando os soros dos animais portadores de outras parasitoses foram testados através do IFI-LVC, reação cruzada foi observada com demodicose e ehrlichiose (18,2 por cento). No EIE-LVC reação cruzada foi observada com um cão portador de ehrlichiose (9,1 por cento). Esses resultados levam as seguintes conclusões: 1) Os resultados obtidos com o kit EIE-LVC indicam que o seu desempenho é similar aquele encontrado no IFI-LVC; 2) Os kits devem ser empregados em associação para obter-se resultados mais seguros. 3) O uso dos kits em associação diminui a possibilidade de ocorrência de resultados falso-positivos e falso-negativos.

Validación de métodos serológicos que se emplean para el diagnóstico de enfermedades reumáticas definidas: Lupus erimatoso sitémico (LES) / Validation of methods serológicos that are used for the diagnosis of defined rheumatic illnesses: Lupus erimatoso sitémico

En el presente trabajo se aplicó un método estadístico no convencional para validar los substratos empleados para detectar anticuerpos antinucleares por inmunofluorescencia indirecta (FANA), comparado con la preuba confirmatoria de ELISA indirecto que mide Anticuerpos anti-dsDNA. Se determinó que los substratos: (línea celular BHK_21 y cortes criostáticos de higado o riñon de rata Ratus novergicus) tenían la misma sensibilidad y valor predictivo negativo (100 por ciento) La línea celular BHK-21 demostró tener mayor especificidad 69 por ciento, valor predictivo positivo 75,8 por ciento, eficiencia 84,3 por ciento y concordancia con los resultados emitidos por la técnica de ELISA (0,81) "muy buena" según el índice de KAppa. Se encontró que el patrón fluorescente difuso a título 1/32 está presente sistemáticamente en todas las muestras tanto positivas comonegativas para anti-ds-DNA. Se demostró que utilizar la línea celular BHK-21 (que reemplaza a la línea tradicional Hep-2), como sustrato "FANA", resulta ser el método taniz, mas eficiente, sensible, específico y de mayor concordancia con los resultados emitidos por un método confirmado para LES, que mide anti-ds-DNA.

Inmunofluorescencia indirecta en un servicio de lactantes de la V Región / Indirect immunofluorescence technique in an infant service, V Region

Este artículo se refiere al examen de inmunofluorescencia indirecta (IFI) utilizado en los servicios de pediatría para el diagnóstico de infecciones respiratorias agudas (IRAS) frecuentes en la infancia; en él se pretende dar cuenta de los aspectos principales a considerar en su toma de muestra, destacando asimismo, el rol fundamental de la enfermera en este procedimiento, teniendo como referente un servicio de lactancia de la V región