Zika Virus Non-Structural Protein 1 Antigen-Capture Immunoassay.

Infection with Zika virus (ZIKV), a member of the Flavivirus genus of the Flaviviridae family, typically results in mild self-limited illness, but severe neurological disease occurs in a limited subset of patients. In contrast, serious outcomes commonly occur in pregnancy that affect the developing fetus, including microcephaly and other major birth defects. The genetic similarity of ZIKV to other widespread flaviviruses, such as dengue virus (DENV), presents a challenge to the development of specific ZIKV diagnostic assays. Nonstructural protein 1 (NS1) is established for use in immunodiagnostic assays for flaviviruses. To address the cross-reactivity of ZIKV NS1 with proteins from other flaviviruses we used site-directed mutagenesis to modify putative epitopes. Goat polyclonal antibodies to variant ZIKV NS1 were affinity-purified to remove antibodies binding to the closely related NS1 protein of DENV. An antigen-capture ELISA configured with the affinity-purified polyclonal antibody showed a linear dynamic range between approximately 500 and 30 ng/mL, with a limit of detection of between 1.95 and 7.8 ng/mL. NS1 proteins from DENV, yellow fever virus, St. Louis encephalitis virus and West Nile virus showed significantly reduced reactivity in the ZIKV antigen-capture ELISA. Refinement of approaches similar to those employed here could lead to development of ZIKV-specific immunoassays suitable for use in areas where infections with related flaviviruses are common.

Improving Immunohistochemistry Capability for Pediatric Cancer Care in the Central American and Caribbean Region: A Report From the AHOPCA Pathology Working Group.

Accessibility to immunohistochemistry (IHC) is invaluable to proper diagnosis and treatment of pediatric patients with malignant neoplasms. Whereas IHC is widely available in anatomic pathology laboratories in high-income countries, access to it in anatomic pathology laboratories of low- and middle-income countries remains a struggle, with many limitations. To advance the quality of the pathology service offered to children with cancer in areas with limited resources, a 5-day pathology training workshop was offered to pathologists and histotechnologists from various countries of the Central American and Caribbean region. An initial assessment of the workshop participants' current laboratory capacities was performed, and a regional training center was selected. Didactic and hands-on activities were offered, and review and evaluation of the IHC slides produced during the training course were compared with original slides from the participants' sites. This model of intensive 5-day training appears to be effective and can potentially be used in other budget-constrained regions. Moreover, it can serve as a continuing education activity for pathologists and histotechnologists, and as part of validations and quality improvement projects to build capacity and develop IHC assay proficiency in low- and middle-income countries.

Efectividad de la reacción en cadena de la polimerasa en tiempo real (PCR-RT) e Inmunodot en el diagnóstico temprano de leptospirosis: análisis comparativo con la prueba de Microaglutinación (MAT) / Effectiveness of the real-time polymerase chain reaction (RT-PCR) and Immunodot in the early diagnosis of Leptospirosis: comparative analysis with the Microagglutination test (MAT)

Introducción: la leptospirosis es una enfermedad infecciosa producida por espiroquetas del género Leptospira. Se disemina a través de la orina de animales domésticos, con mayor frecuencia por roedores. En República Dominicana se necesitan pruebas confiables para el diagnóstico en etapas tempranas de la enfermedad. En el presente trabajo se aborda la validez diagnóstica del PCR en Tiempo Real y del IGM (INMUNODOT) en comparación con la prueba de Microaglutinación (MAT). Materiales y métodos: se realizó un estudio observacional, descriptivo, de corte transversal con 69 pacientes admitidos en el Hospital Regional "José María Cabral y Báez" con diagnóstico presuntivo de leptospirosis desde el 2010 hasta el 2012. Resultados: del total de los casos (69), la mayoría fueron del sexo masculino (94.2 %), menores de 49 años (79.4 %) y provenían de la provincia de Santiago (58 %). La mortalidad fue de 52.1 % de los cuales 52.3 % fueron reportados positivos para Leptospira y 47.6 % resultaron negativos según el MAT. Al comparar los resultados de PCR en relación al MAT se obtuvo una sensibilidad de 27.3 % y una especificidad de 80 %. Los resultados del PCR y del Immunodot fueron equivalentes. Conclusión: en el presente trabajo la realización del PCR en sangre, después del 5to día de inicio de la enfermedad, no demostró ser mejor que la Inmunodot para la detección temprana de la enfermedad, al contrastarlos con el resultado del MAT. En República Dominicana, los casos hospitalizados con diagnóstico presuntivo de leptospirosis permanecen con muy alta mortalidad. Por tanto, es prioritario optimizar el diagnóstico y el tratamiento de estos casos. En este estudio, los casos confirmados con Leptospirosis que fallecieron indican que debe actualizarse el protocolo de tratamiento y asegurarse que pueda implementarse. Por otra parte, los casos no confirmados fallecidos exigen investigar otras causas de enfermedad como la Infección por Hanta Virus.

Introduction: Leptospirosis is an infectious disease caused by spirochaetes of the genus Leptospira. It spread through the urine of domestic animals most frequently in rodents. Need reliable tests to diagnose in early stages of the disease and it has been proposed the use of the PCR in real-time as an option. The present work deals with the diagnostic real-time PCR and IGM (INMUNODOT) in comparison with the Microagglutination (MAT) test. Materials and methods: An observational, descriptive, cross-sectional study with 69 patients admitted as possible Leptospirosis in the Hospital "Jose Maria Cabral y Báez" of Santiago from 2010 to 2012. Results: Of the total cases (69), most were male (94.2%), under 49 years of age (79.4%) coming from the province of Santiago (58%). Mortality was 52.1% of which 52.3% were reported positive for leptospirosis and 47.6% were negative according to the MAT. To compare the results of PCR in relation to the MAT was obtained a sensitivity of 27.3% and a specificity of 80%. The results of the PCR and immunodot were equivalent. Conclusion: In this study, the realization of the PCR in blood after the 5th day of the disease not proved to be better than the Inmunodot for the early detection of the disease. Is necessary to evaluate cases less than 5th day. In the Dominican Republic, hospitalized with a presumptive diagnosis of Leptospirosis cases remain with very high mortality. Therefore, it is important to optimize the diagnosis and treatment of these cases. In this study, confirmed cases with Leptospirosis who died, indicate that you must upgrade the treatment protocol and ensure that it can be implemented. On the other hand, the deceased not confirmed cases require to investigate other causes of disease as the Hanta Virus infection.

Coexpression of major histocompatibility complex class I and LMP1 in Epstein-Barr virus-positive Hodgkin's lymphoma in a pediatric age group.

The Epstein-Barr virus (EBV)-encoded latent membrane proteins (LMP1 and LMP2) are consistently expressed by malignant Hodgkin Reed-Sternberg (HRS) cells of EBV-associated Hodgkin's disease (HD). The EBV LMP1 has been implicated in tumorogenesis. Cytotoxic T lymphocyte (CTL) responses to both of these proteins have been shown in the blood of EBV-seropositive individuals, and in HD, the apparent failure of the CTL response to eliminate HRS cells expressing LMP1 in vivo has given rise to the suggestion that HD may be characterized by the presence of defects in antigen CTL-presentation or in CTL dysfunction. This study examined 28 cases seen at the Regional Children's Hospital Arturo Grullón in Santiago, the Dominican Republic, by immunohistochemistry. High level of expression of major histocompatibility complex (MHC) class I molecules in HRS cells of EBV-associated HD was recorded. These results suggest that deregulation of MHC class I molecules does not account for the apparent ability of EBV to infect HRS cells and to evade CTL protection. Therefore, the result from our work and similar studies will help to determine whether, or which, immunotherapy should be used in positive and negative cases of HD related to EBV.

Condrosarcoma con componente mesequimal adicional

Reportamos el caso de um paciente masculino de 32 años de edad a quien se le diagnosticó condrosarcoma com componente mesenquimal adicional (CCMA). Revisamos la literatura sobre dicha neoplasia haciendo énfasis en el diagnóstico diferencial con otras neoplasias, debido a su importância pronóstica