Prognosis of hospitalized children under 2 years of age with co-detection of influenza A and respiratory syncytial virus at the healthcare facility / Prognóstico de crianças hospitalizadas menores de 2 anos com codetecção de influenza A e vírus sincicial respiratório no local de atendimento

Abstract Objectives: the aim of this study is to evaluate the impact of co-detection of Flu A and RSV using rapid immunochromatographic tests at the point of care, in pediatric patients under 2 years of age in a general hospital. Methods: a retrospective cohort study was conducted to analyze clinical outcomes in hospitalized infants with viral respiratory disease with positive results of rapid immunochromatographic test for RSV and/or Flu-A, from 2013 to 2018. A logistic regression model was adjusted to analyze predictors of orotracheal intubation during hospitalization. Results: we analyzed 220 cases: RSV (192), Flu-A (9), co-detection (19). Lethality rate was 1.8% (2 cases), and 88% (194) were under 1 year of age. Mean time of hospitalizations was higher in patients with co-detection. Variables significantly associated with orotracheal intubation were: younger age in months, comorbidities, RSV and Flu-A co-detection, and bacterial pneumonia during hospitalization. Conclusions: RSV and Flu-Aco-detection was associated with the least favorable clinical prognoses in this study. Rapid test diagnosis may provide important information at the point of care, because molecular panels are not widely accessible in general hospitals. Rapid diagnosis allows timely evaluation and treatment.

Resumo Objetivos: avaliar o impacto da codetecção de Influenza A (FluA) e Vírus Sincicial Respiratório (VSR) por meio de testes imunocromatográficos rápidos em tempo real, em pacientes menores de 2 anos em hospital público e universitário. Métodos: estudo de coorte retrospectivo foi conduzido para analisar os desfechos clínicos de crianças hospitalizadas com doença respiratória viral com resultados positivos do teste rápido imunocromatográfico para VSR e/ou FluA, de 2013 a 2018. Um modelo de regressão logística foi ajustado para analisar preditores de intubação orotraqueal durante a internação. Resultados: foram analisados 220 casos: RSV (192), FluA (9) eco-detecção (19). A letalidade foi de 1,8% (2 casos) e 88% (194) casos em menores de 1 ano. O tempo médio de internação foi maior nos pacientes com codetecção. As variáveis significativamente associadas à intubação orotraqueal foram: menor idade em meses, comorbidades, codetecção de VSR e Flu-A e pneumonia bacteriana durante a internação. Conclusões: codetecção VSR e FluA foi associada a prognósticos clínicos desfavoráveis. O teste rápido fornece informações importantes a beira-leito, pois os painéis moleculares não são amplamente acessíveis em hospitais públicos. O diagnóstico rápido permite a avaliação e tratamento oportunos.

Diagnóstico de cinomose canina por meio de teste imunocromatográfico e sua correlação com achados clínicos e hematológicos no semiárido da Paraíba / Diagnosis of canine distemper by immunochromatographic test and its correlation with clinical and hematological findings in the semiarid region of Paraíba

Objetivou-se através deste trabalho, determinar a prevalência de cinomose canina no semiárido da Paraíba, através de testes rápidos imunocromatográficos, correlacionando-a com os principais achados clínicos e hematológicos. Foram analisadas 67 fichas de animais em que foram realizados testes rápidos para pesquisa de antígeno em amostras nasais e oculares no período de janeiro a dezembro de 2019. Observou-se que 47% (32/67) dos cães analisados foram positivos para cinomose canina. As variáveis que apresentaram diferença estatística significativa (p<0,05) para a infecção foram animais sem raça definida 60% (21/35), animais não vacinados 70% (29/42), e período seco do ano, sendo o mês de agosto (40%; 13/32), com maior ocorrência. Os principais sistemas afetados foram o respiratório 61% (17/28), oftalmológico 70% (22/31), nervoso 69% (13/19), dermatológico 45% (9/20), e gastrintestinal 42% (6/14). As principais alterações hematológicas foram anemia 66% (23/32), leucopenia 76% (19/25) e linfopenia 48% (15/31). Concluiu-se que foi elevada a ocorrência de cinomose canina em animais com suspeita clínica no Semiárido Paraibano, e animais sem raça definida, não vacinados, no período seco do ano foram mais diagnosticados com a enfermidade.

The objective of this study was to determine the prevalence of distemper canine distemper vírus (CDV) infection in the semi-arid region of Paraíba, using rapid immunochromatographic tests, correlating it with the main clinical and hematological findings. 67 records of animals were analyzed in which rapid tests were performed for antigen research in nasal and ocular from January to December 2019. It was observed that 47% (32/67) of compulsory dogs were positive for canine distemper. The variables that defined difference difference (p <0.05) for infection were mixed breed animals 60% (21/35), unvaccinated animals 70% (29/42), and dry period of the year, being the August (40%; 13/32), with greater occurrence. The main affected systems were the respiratory 61% (17/28), ophthalmological 70% (22/31), nervous 69% (13/19), dermatological 45% (9/20), and gastrointestinal 42% (6/14 )) The main changes were hematological, anemia 66% (23/32), leukopenia 76% (19/25) and lymphopenia 48% (15/31). It was concluded that the occurrence of canine distemper in animals with clinical suspicion in the Semiarid Paraibano was high, and non-vaccinated mixed-breed animals in the dry period of the year were more diagnosed with the disease.

Diagnostic parameters and reliability of four rapid immunochromatographic tests for dengue 4

ABSTRACT Background: Although performance of rapid immunochromatographic tests (RITs) for dengue virus (DENV) serotypes 1, 2 and 3 is relatively settled, evidence on accuracy of RITs for DENV-4 are based on studies with small sample sizes and with discrepant results. Objectives: To assess accuracy and inter-observer agreement of RITs targeting dengue nonstructural protein-1 (NS1) antigen - Dengue NS1-Bioeasy™, Dengue NS1 Ag Strip-Bio-Rad™, IVB Dengue Ag NS1-Orangelife™ and Dengue NS1-K130-Bioclin™ in DENV-4 samples. Methods: Study sample (n = 324) included adults presenting at an emergency unit in Rio de Janeiro, Brazil, with fever of ≤72 h and two or more dengue symptoms. A serum sample from each patient was tested by each RIT. A positive reverse-transcription polymerase chain reaction was considered as the reference standard for dengue diagnosis. The diagnostic parameters analyzed for each RIT were sensitivity, specificity, positive and negative predictive values, and likelihood ratios. Each RIT was read by homogeneous (two junior nurses) or heterogeneous (one junior nurse and one senior biologist) pairs. Agreement was estimated by simple kappa with 95% confidence interval, positive (Ppos) and negative (Pneg) proportion concordance and prevalence and bias adjusted kappa, rated from poor (k < 0.0) to almost perfect (0.8 < k < 1.0), and perfect (k = 1). Results: NS1 RITs for DENV-4 diagnosis showed high specificity (95.9%-99.4%), but low sensitivity (14.7%-45.4%). Bioeasy™ had the best performance, with a positive likelihood ratio of 26.0 (95% CI: 8.4;81.0). Inter-observer agreement was almost perfect for all evaluated RITs. Mismatches in confirmed dengue were more common for the Bioclin™ (Ppos 88.3-90.0 %) and Orangelife™ (Ppos 91.7-94.1 %) tests. Conclusions: For DENV-4, the tested RITs had high specificity, but lower sensitivity compared to published results for other serotypes. They should not be used for screening purposes. Different brands may have very different performances. This should be considered upon deciding of using RITs in DENV-4 outbreaks.

Rapid detection of human blood in triatomines (kissing bugs) utilizing a lateral flow immunochromatographic assay - A pilot study

BACKGROUND DNA- and proteomics-based techniques are currently used to identify a triatomine human blood meal. These methods are time consuming, require access to laboratories with sophisticated equipment, and trained personnel. OBJECTIVES We tested a rapid and specific immunochromatographic assay (that detects human blood in forensic samples) to determine if human blood was present in triatomines and their fecal excreta. METHODS We fed Triatoma rubida human blood (positive control) or mouse blood (negative control) and performed the assay on the abdominal contents and fecal excreta. Triatomine field specimens collected in and around human habitations and excreta were also tested. FINDINGS The assay was positive in triatomines fed human blood (N = 5/5) and fecal excreta from bugs known to have ingested human blood (N = 5/5). Bugs feeding on mice (N = 15/15) and their fecal excreta (N = 8/8) were negative for human blood. Human blood was detected in 47% (N = 23/49) triatomines, representing six different species, collected in the field. MAIN CONCLUSIONS The pilot study shows that this rapid and specific test may have applications in triatomine research. Further study is needed to determine the sensitivity of this assay compared to other well-established techniques, such as DNA- and proteomics-based methodologies and the assay's application in the field.

Modification of the Alere GIARDIA Ag TEST immunochromatography KIT methodology for its use in frozen fecal sediment of dogs and cats

ABSTRACT Giardia duodenalis is a worldwide intestinal parasite and is one of the most frequent protozoa species infecting dogs and cats. This study aimed to modify the methodology of Alere GIARDIA Ag TEST KIT for its use in frozen fecal sediments with different storage times in a freezer (-20°C), thus expanding the range of use of this methodology. One hundred fecal sediments from dogs (n=50) and cats (n=50) previously examined by optical microscopy for Giardia cysts were selected for this study. The agreement between the modified immunochromatography and microscopy results was calculated by Kappa coefficient. To evaluate the performance of the modified immunochromatography assay on samples with different storage time, the fecal sediments were divided into three groups according to the time of storage in a freezer: (a) ≤ 1 year (n=37); (b) > 1 year and ≤ 3 years (n=39); (c) > 10 years (max. 13 years) (n=24). The results obtained by the modified immunochromatography assay demonstrates a higher sensitivity of this technique when compared with microscopy, regardless of the frozen storage time. These results allow for the use of this methodology in a greater scope of analysis, especially in frozen fecal sediment triage in sample collections, enabling epidemiological and comparative analysis along different decades.

Evaluación del Aspergillus lateral flow device para el diagnóstico de aspergilosis invasora, experiencia en un hospital universitario / Evaluation of the lateral flow Aspergillus assay for the diagnosis of invasive aspergillosis, experience in a university hospital

Resumen Introducción: El diagnóstico de aspergilosis invasora (AI) se realiza mediante criterios clínicos y microbiológicos los que incluyen marcadores séricos. Recientemente, el test inmunocromatográfico Aspergillus lateral flow device (LFD), ha sido evaluado como método para diagnóstico de AI. Objetivo: Evaluar el desempeño de este test para el diagnóstico de AI. Material y Método: Estudio transversal en que se evaluaron muestras de suero y lavado bronco-alveolar (LBA) procesadas para galactomanano provenientes de pacientes adultos con sospecha de AI, atendidos en el Hospital Clínico de Red de Salud UCCHRISTUS. Resultados: Se procesó un total de 142 muestras de 98 pacientes, correspondientes a AI probada 5,6%, AI probable 41,5%, AI posible 12,7% y ausencia de AI 40,1%. Al confrontar los resultados con las categorías diagnósticas según criterios EORTC/MSG se obtuvo una sensibilidad y especificidad de LFD para diagnóstico de AI de 70,9 y 53,5% para muestras de suero y 83,3 y 38,5% para muestras de LBA. La concordancia entre galactomanano y LFD fue de 62,4% (54,1-69,9) con un índice Kappa de 0,202 (0,03682-0,3669). Conclusiones: Aspergillus LFD presentó una adecuada sensibilidad; sin embargo, la especificidad fue baja por lo que un resultado positivo requiere ser confirmado.

Background: The incidence of invasive aspergillosis is increasing. Its diagnosis is based on clinical and microbiological criteria which include the determination of serological markers such as galactomannan. Recently, the Aspergillus lateral flow device, an inmunocromatograph assay has been described for its diagnosis. Aim: To evaluate the performance of the lateral flow device for the diagnosis of invasive aspergillosis (IA) in adult patients. Material and Method: In this cross-sectional study, frozen samples that had been previously evaluated for galactomannan from patients classified with proven/probable/possible or no AI according to the EORTC/MSG criteria were selected. Results: A total of 142 samples from 98 patients were processed, corresponding to proven AI 5.6%, probable IA 41.5%, possible IA 12.7% and no-IA 40.1%. The sensitivity and specificity of the Aspergillus lateral flow was 70.9% and 53.5% for serum samples and 83.3% and 38.5% for BAL samples. The concordance between the galactomannan and Aspergillus lateral flow was 62.4% (54.1 - 69.9) with a Kappa index of 0.202 (0.03682 - 0.3669). Conclusions: We observed a good sensitivity but low specificity, a positive result need a confirmatory test.

Development of a prototype immunochromatographic test for rapid diagnosis of respiratory adenovirus infection

Abstract Human adenoviruses comprise an important group of etiologic agents that are responsible for various diseases in adults and children, such as respiratory, ocular, gastroenteric, and urinary infections. In immunocompromised and organ-transplanted individuals, these agents can cause generalized infections. Rapid diagnostic methods for detecting these infectious agents are not widely available.The aim of this work was to produce monoclonal and polyclonal anti-adenovirus antibodies to be used in a rapid diagnostic test for respiratory infections.Adenovirus hexons were satisfactorily purified by ultracentrifugation and chromatography. After virus purification, anti-hexon monoclonal antibodies were produced and characterized, following classical methods. Antibodies were specific for adenoviruses 2, 3, 5, and 41. The proposed immunochromatographic test was standardized using colloidal gold.The standardization of the rapid test was sufficient to detect adenovirus antigens (in nasopharyngeal lavage samples) with sensitivity of 100% and specificity of 85% when compared to direct immunofluorescence.The immunochromatographic assay prototype was sufficiently sensitive to detect B (3), C (2 and 5), and F (41) adenovirus samples. Although based on preliminary data, the test demonstrated the same performance as direct immunofluorescence, but with the advantage of being a point-of-care test. Further studies are still needed to confirm its effectiveness in clinical practice.

Análisis de técnicas treponémicas y no treponémicas en el tamizaje serológico de sífilis / Analysis of treponemical and non treponemical techniques for syphilis blood bank screening

Introducción: las infecciones producidas por el Treponema pallidum causante de la sífilis han alcanzado gran trascendencia entre las enfermedades infecciosas transmitidas por transfusión (ITT) y un nuevo repunte a nivel mundial. Debido a las diferentes etapas clínicas que presenta la enfermedad, el desempeño de cada prueba de detcción varía y se limita. Así, la elección de una técnica de tamizaje adecuada se convierte en un punto fundamental para garantizar la calidad y seguridad de cada hemocomponente despachado. Objetivo: analizar y evaluar la eficacia de cuatro técnicas de tamizaje serológico. Métodos: se realizó un estudio comparativo transversal con 1 376 muestras seleccionadas al azar a nivel nacional en el mes de diciembre 2015. Se compararon las técnicas de inmunocromatografía (IC), floculación (VDRL), electroquimioluminiscencia (ECLIA) y microelisa. Se analizó la eficacia individual de cada técnica y comparativa con respecto a la gold standard (FTA-ABS) utilizando para ello el coeficiente de correlación Cohen-Kappa (К). Resultados: las cuatro pruebas presentaron un nivel de concordancia del 98,67 por ciento. Del total de resultados discrepantes el 63,16 por ciento fueron generados por VDRL, la cual al mismo tiempo demostró tener el menor rendimiento (k=0,14) y alcanzó los valores más bajos de sensibilidad (s=69 por ciento) y especificidad (e=45 por ciento), lo cual contrastó con la IC que demostró el mayor rendimiento (k=0,863, s=100 por ciento, e=0,8 2 por ciento), seguido de la ECLIA (k=0,801, s=96 por ciento, e=0,82 por ciento) y el microelisa (k=0,711, s=100 por ciento, e=0,64 por ciento). Conclusión: se evidencia la necesidad de utilizar pruebas de nuevas tecnologías en el tamizaje serológico de sífilis y remplazar el uso de VDRL, ya que una correcta selección asegura el descarte de hemocomponentes en el número correcto (evitando grandes pérdidas de sangre y de dinero) y, en especial se asegura la calidad sanitaria de cada hemocomponente(AU)

Introduction: Infections produced by Treponema pallidum, which causes syphilis, have reached an important high level among infectious transmitted by transfusion (ITT). Due to the different clinical stages of the disease, the performance of each test is varied and limited. Thus, the choice of a suitable screening technique becomes a fundamental point in a blood bank, to guarantee the quality and safety of each blood component dispatched. Objective: The aim of this study was to analyze and evaluate the performance of four serological screening techniques. Methods: A cross-sectional study was conducted with 1 376 randomly selected samples nationwide in December 2015. The techniques used were immunochromatography (IC), flocculation (VDRL), electrochemiluminescence (ECLIA) and microelisa, and we compared the performance of each one and with respect to the gold standard (FTA-ABS) by using the Cohen-Kappa correlation coefficient (K). Results: The four tests had a concordance level of 98.67 percent. Of the total discrepant results the 63.16 percent were generated by VDRL, which at the same time showed the worst performance (k=0,14) and reached the lowest values ​​of sensitivity (s=69 percent) and specificity (e=45 percent). That contrasted with IC, which showed the best performance (k=0,883, s=100 percent, e=82 percent), followed by ECLIA (k=0,801, s=96 percent, e=0,82 percent) and microelisa (k=0,711, s=100 percent, e=0.64 percent). Conclusion: There is a necessity to use tests with new technologies in the serological screening of syphilis and to replace the use of VDRL in a blood bank, due to a correct selection, ensures the quality and the disposal of blood components in the correct number avoiding great losses of blood and money(AU)

Displacers improve the selectivity of phosphopeptide enrichment by metal oxide affinity chromatography / Uso de desplazadores para mejorar la selectividad de los fosfopéptidos enriquecidos por cromatografía de afinidad con óxidos de metales

Abstract: Background: A key process in cell regulation is protein phosphorylation, which is catalyzed by protein kinases and phosphatases. However, phosphoproteomics studies are difficult because of the complexity of protein phosphorylation and the number of phosphorylation sites. Methods: We describe an efficient approach analyzing phosphopeptides in single, separated protein by two-dimensional gel electrophoresis. In this method, a titanium oxide (TiO2)-packed NuTip is used as a phosphopeptide trap, together with displacers as lactic acid in the loading buffer to increase the efficiency of the interaction between TiO2 and phosphorylated peptides. Results: The results were obtained from the comparison of mass spectra of proteolytic peptides of proteins with a matrix-assisted laser desorption-ionization-time of flight (MALDI-TOF) instrument. Conclusions: This method has been applied to identifying phosphoproteins involved in the symbiosis Rhizobium etli-Phaseolus vulgaris.

Resumen: Introducción: Un proceso clave en la regulación celular es la fosforilación de proteínas, que se lleva a cabo por cinasas y fosfatasas. Sin embargo, los estudios de fosfoproteómica son difíciles debido a la complejidad de la fosforilación proteica y el número de sitios de fosforilación. Métodos: En el presente trabajo se describe una eficiente estrategia metodológica para analizar fosfopéptidos de proteínas separadas mediante electroforesis bidimensional. En este método, una columna con microesferas de dióxido de titanio (TiO2/NuTip) se utilizó para atrapar los fosfopéptidos en la superficie del TiO2 previamente empacado en una punta. El uso de desplazadores en el buffer de carga, como el ácido láctico, mejoró significativamente la selectividad. Resultados: Los resultados se obtuvieron mediante la comparación de los espectros de masas de péptidos proteolíticos de proteínas analizados utilizando un instrumento de desorción/ionización láser asistida por matriz-tiempo de vuelo (MALDI-TOF). Conclusiones: Este método se ha aplicado para la identificación de fosfoproteínas involucradas en la simbiosis del Rhizobium etli con Phaseolus vulgaris.

Detección de Brucella spp. por un sistema inmunocromatográfico comercial, en muestras ambientales cubanas / Detection of Brucella spp using a commercial system in enviroment cuban samples

Introducción: actualmente en Cuba se desconoce la circulación de las especies de Brucella en el medio ambiente, por la inexistencia de métodos de laboratorio que permitan su identificación. Objetivos: detectar Brucella spp. en muestras ambientales cubanas aplicando un sistema inmunocromatográfico comercial. Métodos: se estudiaron 59 muestras ambientales de una zona endémica de brucelosis bovina y 50 muestras ambientales de zonas controladas de la enfermedad, durante el período diciembre de 2011 y hasta febrero de 2012. Se utilizó el sistema inmunocromatográfico directo de flujo lateral para Brucella spp. comercializado en Cuba. Resultados: el 52,5 por ciento (31/59) de las muestras ambientales de la zona endémica resultaron positivas para Brucella spp. Las muestras ambientales que presentaron el mayor porcentaje de positividad fueron el estiércol (62,5 por ciento ) y las del suelo cementado (26,9 por ciento ). Predominaron las reacciones fuertemente positivas en un 74,1 por ciento (23/31). Conclusiones: el sistema inmunocromatográfico comercial detecta un elevado porcentaje de Brucella spp. en muestras ambientales de la zona endémica cubana, lo que pudiera avalar su implementación en la red de laboratorios de Salud Pública y del Instituto de Medicina Veterinaria de Cuba. Los resultados de esta investigación deben complementarse con los aislamientos de brucelas en el medio ambiente, pacientes humanos y en animales(AU)

Introduction: Information about the circulation of Brucella species in the Cuban environment is not currently available due to the lack of laboratory methods for their identification. Objectives: Detect Brucella spp. in Cuban environmental samples using a commercial immunochromatographic system. Methods: A study was conducted of 59 environmental samples from an endemic zone for bovine brucellosis and 50 environmental samples from areas where the disease has been controlled, from December 2011 to February 2012. Use was made of the direct lateral flow immunochromatographic system for Brucella spp. commercially available in Cuba. Results: Of the environmental samples from the endemic zone 52.5 percent (31/59) tested positive for Brucella spp. The environmental samples showing the highest positivity were those taken from dung (62.5 percent) and cemented soil (26.9 percent). Strongly positive reactions predominated in 74.1 percent (23/31) of the samples. Conclusions: The commercial immunochromatographic system used detected a high percentage of Brucella spp. in environmental samples from the endemic zone, which could justify its implementation in the Public Health laboratory network and the Institute of Veterinary Medicine of Cuba. The results obtained should be complemented with Brucella isolates from the environment, human patients and animals(AU)

The Usefulness of a Novel Screening Kit for Colorectal Cancer Using the Immunochromatographic Fecal Tumor M2 Pyruvate Kinase Test

BACKGROUND/AIMS: M2 pyruvate kinase (M2-PK) is an enzyme that is produced in undifferentiated and proliferating tissues. This study aims to evaluate the usefulness of the immunochromatographic M2 pyruvate kinase (iM2-PK) for the screening of colorectal cancer (CRC) and premalignant lesions. METHODS: Healthy volunteers and patients with colorectal neoplasia were enrolled in six academic hospitals in the capital province of Korea. The iM2-PK value was compared with the immunochromatographic fecal occult blood test (iFOBT) and fecal tumor M2-PK enzyme-linked immunosorbent assay (ELISA). RESULTS: A total of 323 subjects were enrolled. The sensitivity of iM2-PK for CRC was 92.8%, which was superior to iFOBT (47.5%, p<0.0001). For adenomatous lesions, the sensitivity of iM2-PK was 69.4%, which was also superior to iFOBT (12.1%, p<0.001). Compared with M2-PK ELISA, iM2-PK exhibited significantly enhanced sensitivity for CRC (97.5% vs 80.0%, p=0.0289). The sensitivity of iM2-PK was higher in advanced stages of CRC compared with cancers confined to the mucosa and submucosa (p<0.05). However, lymph node metastasis had no influence on the sensitivity of iM2-PK. CONCLUSIONS: The iM2-PK exhibited increased sensitivity for identifying CRC and adenomatous lesions compared with iFOBT. Given its rapid results and convenience, CRC screening using iM2-PK is promising.

Performance of the SD Bioline TB Ag MPT64 Rapid test for quick confirmation of Mycobacterium bovis isolates from animals

Mycobacterium (M.) bovis, a bacterium in the M. tuberculosis complex, is a causative agent of bovine tuberculosis, a contagious disease of animals. Mycobacterial culture is the gold standard for diagnosing bovine tuberculosis, but this technique is laborious and time-consuming. In the present study, performance of the SD Bioline TB Ag MPT4 Rapid test, an immunochromatographic assay, was evaluated using reference bacterial strains and M. bovis field isolates collected from animals. The SD MPT64 Rapid test produced positive results for 95.5% (63/66) of the M. bovis isolates from cattle and 97.9% (46/47) of the isolates from deer. Additionally, the test had a sensitivity of 96.5% (95% CI, 91.2-99.0), specificity of 100% (95% CI, 96.7-100.0), positive predictive value of 100% (95% CI, 96.7-100.0), and negative predictive value of 92.9% (95% CI, 82.7-98.0) for M. bovis isolates. In conclusion, the SD MPT64 Rapid test is simple to use and may be useful for quickly confirming the presence of M. bovis in animals.

Associação de hipovitaminose D com Lúpus Eritematoso Sistêmico e inflamação / Association of hypovitaminosis D with Systemic Lupus Erythematosus and inflammation

Introdução: Atualmente, é descrita elevada prevalência de hipovitaminose D no Lúpus Eritematoso Sistêmico (LES), a qual se associa a algumas manifestações clínicas e maior atividade inflamatória. Objetivo: Avaliar a associação entre insuficiência de vitamina D com LES e marcadores inflamatórios. Métodos: Estudo transversal, tendo sido avaliados 45 pacientes com LES e 24 controles sem a doença. Níveis de 25-hidroxivitamina D [25(OH)D] menores que 30 ng/mL foram considerados insuficientes. A atividade da doença foi avaliada pelo Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). Foram avaliados, ainda, proteína C reativa ultrassensível (PCRus) e interleucina-6 (IL-6) para verificação do status inflamatório. Para avaliação do envolvimento renal, foram realizados análise de elementos anormais e sedimentoscopia urinárias (EAS), hematúria e piúria quantitativas, proteinúria e depuração de creatinina em urina de 24 horas e anti-DNA de dupla hélice sérico. Resultados: A prevalência de insuficiência de 25(OH)D foi de 55% nos pacientes lúpicos e 8% nos participantes controles (p = 0,001). A mediana da 25(OH)D foi menor nos pacientes do que no grupo controle. Os pacientes com insuficiência de 25(OH)D apresentaram níveis mais elevados de IL-6 e maior prevalência de hematúria ao EAS. Não houve correlação entre vitamina D, nefrite lúpica e SLEDAI. Conclusão: Em nosso estudo, a insuficiência de vitamina D foi mais prevalente em pacientes com LES e se associou com níveis mais elevados de IL-6 e presença de hematúria. .

Introduction: Nowadays it is described a high prevalence of hypovitaminosis D in Systemic Lupus Erythematosus (SLE), which is associated with some clinical manifestations and increased inflammatory activity. Objective: To evaluate the association between vitamin D insufficiency with SLE and inflammatory markers. Methods: Cross-sectional study, in which have been evaluated 45 SLE patients and 24 controls without the disease. Levels of 25-hydroxyvitamin D [25(OH) D] less than 30 ng/mL were considered inadequate. Disease activity was assessed by the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). High sensitivity C reactive protein (hsCRP) and interleukin-6 (IL-6) were evaluated for verification of the inflammatory status. For assessment of renal involvement, analysis of abnormal elements and urinay sediment (AES), quantitative hematuria and pyuria, proteinuria and creatinine clearance in 24-hour urine and serum anti-double stranded DNA were performed. Results: The prevalence of 25(OH)D insufficiency was 55% in SLE patients and 8% in the controls participants (p = 0.001). The median of 25(OH)D was lower in patients than in controls. Patients with insufficient 25(OH)D had higher levels of IL-6 and higher prevalence of hematuria in the AES. There was no correlation between vitamin D and SLEDAI or lupus nephritis. Conclusion: In our study, vitamin D deficiency was more prevalent in patients with SLE and was associated with higher levels of IL-6 and hematuria. .

Evaluación de la técnica BD MGITTM TBc® para identificación del complejo Mycobacterium tuberculosis / Evaluation of the BD MGITTM technique for identifying the Mycobacterium tuberculosis complex

Objetivo Evaluar la técnica BD MGITTM TBc® para identificación del complejo Mycobacterium tuberculosis a partir de aislamientos en medio de cultivo sólido y líquido. Materiales y Métodos Se desarrolló un estudio descriptivo, donde se analizaron 117 aislamientos por la técnica de inmucromatografía a partir de cultivos en medio sólido y líquido para identificación del complejo Mycobacterium tuberculosis. Se calculó coeficiente kappa para determinar el grado de acuerdo entre los dos métodos. Cuando hubo diferencia de resultados estos se confirmaron mediante pruebas convencionales. La herramienta empleada para el análisis de datos fue Epidat 3.1. Resultados La metodología BD MGITTM TBc® realizada a partir de cultivos en medio sólido y líquido, presentó un grado de acuerdo excelente con un coeficiente kappa de 0,84. Conclusión La técnica BD MGITTM TBc® realizada a partir de cultivos en medio sólido, para la identificación del complejo Mycobacterium tuberculosis, presenta excelente concordancia, comparada con los resultados obtenidos en medio de cultivo líquido. El Laboratorio Nacional de Referencia recomienda el uso de esta técnica para la identificación de especie en medio de cultivos sólidos.(AU)

Objetive To evaluate BD MGITTM TBc® technique for identifying the Mycobacterium tuberculosis complex using isolates obtained in liquid and solid media. Methods A descriptive study was conducted in which 117 isolates were analyzed by the immune-chromatography technique obtained from solid and liquid cultures to identify the Mycobacterium tuberculosis complex. The kappa coefficient was calculated to determine the degree of agreement between the two methods .When there were different results, they were confirmed with a conventional test. The tool used to analyze the data was Epidat 3.1 Results The BD MGITTM TBc® methodology performed in solid and liquid culture isolates, showed an excellent degree of agreement with a kappa coefficient 0.84. Conclusion The BD MGITTM TBc® technique using solid media culture isolates for the identification of the Mycobacterium tuberculosis complex has a good correlation compared to results obtained from liquid media culture isolates. The Reference National Laboratory recommends the use of this technique for the identification of species in solid media culture isolates.(AU)

Comparison of seven diagnostic tests to detect Trypanosoma cruzi infection in patients in chronic phase of Chagas disease / Comparación de siete pruebas diagnósticas para detectar infección por Trypanosoma cruzi en pacientes en fase crónica de la enfermedad de Chagas

Objective: To compare the diagnostic performance of seven methods to determine Trypanosoma cruzi infection in patients with chronic Chagas disease. Methods: Analytical study, using the case-control design, which included 205 people (patients with Chagasic cardiomyopathy, n= 100; control group, n= 105). Three enzyme linked immunosorbent assays, one indirect hemagglutination assay and one immunochromatographic test were assessed. Additionally, DNA amplification was performed via the PCR method using kinetoplast and nuclear DNA as target sequences. For the comparative analysis of diagnostic tests, the parameters used were sensitivity, specificity, positive and negative predictive values, Receiver Operator Characteristic (ROC), positive and negative likelihood ratio, as well as κ quality analysis. Results: The commercial Bioelisa Chagas test showed the highest sensitivity (98%), specificity (100%), and positive and negative predictive values; additionally it had the highest discriminatory power. Otherwise, the amplification of T. cruzi DNA in blood samples showed low values of sensitivity (kinetoplast DNA= 51%, nuclear DNA= 22%), but high values of specificity (100%), and moderate to low discriminatory ability. Conclusion: The comparative analysis among the different methods suggests that the diagnostic strategy of T. cruzi infection in patients with chronic Chagas disease can be performed using ELISA assays based on recombinant proteins and/or synthetic peptides, which show higher diagnosis performance and can confirm and exclude the diagnosis of T. cruzi infection. The molecular methods show poor performance when used in the diagnosis of patients with chronic Chagas disease.

Objetivo: Comparar la capacidad diagnóstica de siete métodos para determinar infección por Trypanosoma cruzi, en pacientes con enfermedad de Chagas crónica. Métodos: Estudio analítico de casos y controles, que incluyó 205 personas (pacientes con miocardiopatía chagásica, n= 100; grupo control, n= 105). Se evaluaron tres inmunoensayos enzimáticos, una hemaglutinación indirecta y una inmunocromatografia. Adicionalmente, se realizó amplificación de ADN de T. cruzi por reacción en cadena de la polimerasa utilizando como secuencias diana ADN de kinetoplasto y nuclear. Para el análisis comparativo de las pruebas diagnósticas, los parámetros utilizados fueron sensibilidad, especificidad, valores predictivo positivo y negativo, análisis ROC, razón de verosimilitud positiva y negativa, así como análisis de calidad κ. Resultados: La prueba Bioelisa para Chagas mostró la mayor sensibilidad (98%), especificidad (100%) y valores predictivos positivo y negativo; además ésta tuvo el mayor poder discriminatorio. En contraste, los ensayos de amplificación de ADN de T. cruzi mostraron baja sensibilidad (ADN de kinetoplasto= 51%, ADN nuclear= 22%), alta especificidad (100%) y de moderada a baja capacidad discriminatoria. Conclusión: El análisis comparativo entre los métodos sugiere utilizar como estrategia diagnóstica en pacientes crónicos con enfermedad de Chagas, los ensayos de ELISA con proteínas recombinantes y/o péptidos sintéticos por mostrar un rendimiento diagnóstico superior y tener la capacidad de confirmar y descartar el diagnóstico de infección por T. cruzi. Los métodos moleculares muestran pobre rendimiento para ser utilizados en el diagnóstico de pacientes en fase crónica con enfermedad de Chagas.

Comparison of Two Rapid Immunochromatographic Assays (ICT Malaria P.f. /P.v. Test and OptiMAL Test) with Microscopy for Detection of Malaria Parasites.

This study was done to compare the ability of newly developed immunochromatographic assays (ICT), i.e., ICT malaria P.f. / P.v. test and optiMAL test with standard microscopy for the diagnosis of malaria. ICT P.f. / P.v. test detects Plasmodium falciparum specific histidine rich protein-2 (HRP2) antigen and a pan-malarial common specific antigen, where as optiMAL test detects P. falciparum specific parasite Lactate Dehydrogenase (pLDH) enzyme and a common specific pLDH enzyme. Material and Methods: Blood samples were obtained from 150 patients clinically diagnosed as malaria between July 2011 to December 2011.The venous blood were tested for malaria by microscopy and simultaneously ICT P.f./P.v.and optiMAL tests. Results: From total 150 samples, 59 (39.3%) were positive by blood films while 64 (42.7%) were positive by ICT p.f. / p.v. and 52 (34.7%) by optiMAL tests. The blood film indicated that 32.2% (19 of 59) of patients were positive for P. vivax and 67.8% (40 of 59) were infected with P. falciparum. ICT P.f./P.v. test showed 23.4% (15 of 64) were positive for P. vivax and 76.6% (49 of 64) were infected with P. falciparum. Similarly, optiMAL test detected 30.8% (16 of 52) were positive for P. vivax and 69.2% (36 of 52) were infected with P. falciparum. ICT P.f./P.v. test had sensitivities 78.9%, 87.5% and specificities 100%, 87.3% for P. vivax and P. falciparum respectively. optiMAL test showed sensitivities 84.2%, 80% and specificities 100%, 96.4% for P. vivax and P. falciparum respectively. Conclusion: These rapid immunoassays (ICT P.f./P.v. and optiMAL) tests can be used as supplementary to traditional light microscopy for the diagnosis of malarial parasites.

Purification of a protease inhibitor from Dolichos biflorus using immobilized metal affinity chromatography.

Plant protease inhibitors (PIs) are generally small proteins which play key roles in regulation of endogenous proteases and may exhibit antifeedant, antifungal, antitumor and cytokine inducing activities. Dolichos biflorus (horse gram) is an unexploited legume, which is rich in nutrients and also has therapeutic importance. It contains a double-headed PI, which is an anti-nutritional factor. As there is no report available on its simultaneous removal and purification in single step, in this study, a double-headed PI active against both trypsin and chymotrypsin was purified from Dolichos biflorus to ~14-fold with ~84% recovery using an immobilized metal affinity chromatography (IMAC) medium consisting of Zn-alginate beads. The method was single-step, fast, simple, reliable and economical. The purified inhibitor showed a single band on SDS-PAGE corresponding to molecular mass of 16 kDa and was stable over a pH range of 2.0-12.0 and up to a temperature of 100°C for 20 min. The optimum temperature for trypsin and chymotrypsin inhibitor was observed to be 50°C and 37°C, respectively and pH optimum was pH 7.0 and 8.0, respectively. Thus, IMAC using Zn-alginate beads was useful in simultaneous purification and removal of an anti-nutritional factor from horse gram flour in single step. This procedure may also be employed for purification of other plant PIs in one step.

Epidemiología de la enfermedad de Chagas en menores en riesgo de la provincia de Santa Fe y evaluación de una nueva técnica diagnóstica en terreno / Epidemiology of Chagas disease in children at risk in the province of Santa Fe and evaluation of a new diagnostic technique in field

La Enfermedad de Chagas (ECh) es una endemia cuya transmisión vectorial ha ido disminuyendo en Latinoamérica. No obstante el riesgo de transmisión por esta vía sigue siendo importante en algunas regiones, entre ellas la Eco Región del Gran Chaco, debido a factores naturales y a fuertes condicionantes sociales: población rural, aislada y con escaso peso económico y político. En Santa Fe estas condiciones se dan en los distritos rurales de los Departamentos 9 de Julio y Vera, en el norte de la Provincia. Los análisis serológicos en terreno tienen importancia tanto para la salud individual como colectiva. La realización de estos análisis se ve dificultada por la distancia a los laboratorios con recursos humanos e instrumentales adecuados. En ese sentido, las técnicas inmunocromatográficas (IC) son promisorias por cuanto no requieren de instrumental de laboratorio ni personal especializado. Una IC para diagnóstico de ECh recientemente desarrollada fue probada por el fabricante en un número reducido de muestras de paneles de sueros estándar y en condiciones de laboratorio, mostrando buena sensibilidad (S) (>95%) y especificidad (E) (>97%). Sin embargo es desconocida su performance con sangre entera y en condiciones de terreno. Los objetivos de la presente tesis fueron realizar una descripción epidemiológica de la ECh en niños de 5 a 14 años de distritos rurales del Chaco Santafesino y evaluar el desempeño de una IC en terreno...

The Chagas Disease (ChD) is a vector-borne endemic disease which has been declining in Latin America. However the risk of transmission by this route is still important in some regions, including the Gran Chaco Eco Region, due to natural factors and tough social conditions: a rural, isolated population with a low economic and political influence. In Santa Fe these conditions exist in the rural districts of the departments of 9 de Julio and Vera, in the north of the province. Serological analyses on field are important for both the individual and public health. The accomplishment of these analyzes is hampered by the distance to laboratories with adequate human and instrumental resources. In that sense, the immunochromatographic techniques (IC) are promising because they do not require laboratory instruments or specialized personnel. A newly developed IC for the diagnostic of Chd was tested by the manufacturer in a small sample of standard sera panels and in laboratory conditions, showing good sensitivity (S) (> 95%) and specificity (Sp) (> 97% ). However its performance is unknown with whole blood and in the field conditions. The objectives of this thesis were to conduct an epidemiological description of the ChD in children of 5-14 years old from the Chaco Santafesino rural districts and evaluate the performance of an IC in the field...

Avaliação epidemiológica de doenças negligenciadas em escolares: filariose linfática e parasitoses intestinais / Epidemiological assessment of neglected diseases in children: lymphatic filariasis and soil-transmitted helminthiasis

OBJETIVO: Descrever a prevalência de infecção filarial e de parasitoses intestinais em escolares numa área endêmica de filariose e refletir sobre a opção terapêutica utilizada no Brasil no tratamento coletivo para filariose. MÉTODOS: Estudo transversal envolvendo 508 alunos na faixa etária de 5-18 anos cadastrados em escolas públicas do município de Olinda-PE. Realizou-se a investigação da parasitose intestinal em três amostras de fezes, analisadas pelo método de Hoffmann, Pons e Janer. A investigação filarial foi feita com teste antigênico pela técnica de imunocromatográfica rápida (ICT) e pesquisa de microfilárias, utilizando filtração em membrana de policarbonato. Para análise de dados utilizou-se a estatística descritiva através do programa EpiInfo versão 7. RESULTADOS: A prevalência de filariose por ICT foi de 13,8% e por microfilaremia de 1,2%, enquanto a de parasitoses intestinais foi 64,2%. A concomitância do diagnóstico filarial e de parasitoses intestinais foi de 9,4% e, 31,5% eram isentos de ambas as parasitoses. Entre os indivíduos com parasitoses intestinais, 55% eram monoparasitados e 45% poliparasitados. A presença de geohelmintos ocorreu em 72,5% dos parasitados. No grupo com infecção filarial a ocorrência de geohelmintíase foi de 54,5%. CONCLUSÕES: O diagnóstico simultâneo de infecção filarial e parasitose intestinal, bem como a elevada frequência de geohelmintos justificam uma reavaliação da estratégia terapêutica do tratamento coletivo no programa de filariose no Brasil.

OBJECTIVE: To report the prevalence of lymphatic filariasis and intestinal parasitic infections in school-aged children living in a filariasis endemic area and discuss about the therapeutic regimen adopted in Brazil for the large-scale treatment of filariasis. METHODS: A cross-sectional study including 508 students aged 5-18 years old, enrolled in public schools within the city of Olinda, Pernambuco. The presence of intestinal parasites was analyzed using the Hoffman, Pons and Janer method on 3 stool samples. The diagnosis of filarial infection was performed using the rapid immunochromatographic technique (ICT) for the antigen, and the polycarbonate membrane filtration for the presence of microfilariae. Descriptive statistics of the data was performed using EpiInfo version 7. RESULTS: The prevalence of filariasis was 13.8% by ICT and 1.2% by microfilaraemia, while intestinal parasites were detected in 64.2% of cases. Concurrent diagnosis of filariasis and intestinal parasites was 9.4%, while 31.5% of students were parasite-free. Among individuals with intestinal parasites, 55% had one parasite and 45% had more than one parasite. Geohelminths occurred in 72.5% of the parasited individuals. In the group with filarial infection the prevalence of soil-transmitted helminthiasis was 54.5%. CONCLUSIONS: The simultaneous diagnosis of filariasis and intestinal parasites as well as the high frequency of geohelminths justify the need to reevaluate the treatment strategy used in the Brazilian filariasis large-scale treatment program.