Programa de Pós-graduação em Medicina Tropical e Saúde Pública
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Item Avaliação de diferentes métodos para a classificação de pacientes e de carreadores de antígenos empregados na sorologia de hanseníase(Universidade Federal de Goiás, 2014-03-31) Moura, Rodrigo Scaliante de; Stefani, Mariane Martins de Araújo; Bührer, Samira; http://lattes.cnpq.br/6715236625981869; Buhrer, Samira; Penna, Gerson de Oliveira; Grossi, Maria Aparecida de Faria; Kipnis, Ana Paula; Silveira, Lucimeire Antonelli daSerology to detect IgM antibodies against the PGL-I, a species-specific antigen from Mycobacterium leprae, can be used to aid in the identification of individuals with high bacillary load. Samples and the database from the U-MDT clinical trial were used to compare and report the perfomance of diverse classification methods. This study also evaluated the possibility of enhancement of serological tests by substituting the antigen carrier protein, the bovine serum albumin (BSA) by a human protein (HSA), aiming to eliminate unespecific bindings. The tests specificity was evaluated using sampes from household contacts, healthy population, TB patients and Visceral Leishmaniasis from endemic and non-endemic areas. In this study the most sensitive and the least specific approach was represented by counting the number of skin lesions, which identifiel 99% of MB leprosy patients. ML Flow HSA tests correcty allocated 70.9% and 68.6% of patients in the PB group, and 87% and 81% of patients in the MB group, respectively. The study suggests that the carrier protein of the antigen did not influence significantly in the seropositivity of studied groups. PGL-I serology did not presented cross-reactions with Visceral Leishmaniasis serum samples and confirmed a low positivity among control groups.Item Otimização e produção do teste rápido para controle da hanseníase (ML Flow)(Universidade Federal de Goiás, 2011-03-01) Moura, Rodrigo Scaliante de; Stefani, Mariane Martins de Araújo; Bührer, Samira; http://lattes.cnpq.br/6715236625981869; Buhrer, Samira; Grossi, Maria Aparecida de Faria; Kipnis, Ana Paula Junqueira(não consta)Item Desenvolvimento de teste rápido para detecção de Listeria monocytogenes em alimentos(Universidade Federal de Goiás, 2018-02-27) Silva Filho, Ernandes da; Kipnis, André; http://lattes.cnpq.br/4434965360286741; Bührer, Samira; http://lattes.cnpq.br/6715236625981869; Bührer, Samira; Souza, Guilherme Rocha Lino de; Moura, Rodrigo Scaliante de; André, Maria Cláudia Dantas Porfírio Borges; Pinto, Emerith Mayra HungriaListeriosis is a bacterial infection caused by Listeria monocytogenes, a bacterium transmitted mainly by the ingestion of contaminated food. It is the main responsible for food infections with low morbidity rate, but when contamination occurs in immunosuppressed the mortality rates are high, representing an important public health problem. Methods available for identification of the presence of Listeria monocytogenes include isolation of the bacterium through conventional culture, laborious, time-consuming and low sensitivity method, and usually complex and costly molecular and / or immunological methods requiring trained personnel and equipment. Therefore, the search for rapid, sensitive and specific methods for detecting the presence of Listeria monocytogenes in foods is useful and necessary for infection prevention and great value to reduce the mortality rate in the immunocompromised as well as to enable the epidemiological surveillance of this disease. The objectives of the study were: To develop lateral flow immunochromatographic test for the identification of L. monocytogenes in foods; To evaluate the performance of the lateral flow immunochromatographic test using food samples naturally and artificially contaminated with L. monocytogenes. As the detection agent, were impregnated glass fiber with anti-internalin-A MAb-2D12 conjugated to colloidal gold. As a capture agent, were sensitized, on nitrocellulose paper, the test line with polyclonal anti-internalin-B antibody, and in the control line with protein A, to validate the activity of the detection agent. Inoculations of standard strains of Listeria monocytogenes were used for evaluation of the Listeria detection in the proposed test. As a negative control, we used strains of Listeria innocua, Enterobacter sp. and Bacillus cereus. Positivity was obtained in prototypes of rapid tests sensitized with Polyclonal Anti-InlB. Positivity was observed in samples containing culture medium and newly cultured bacteria, validating the methodology used in the development of the test. The detection capacity of the test at D concentration is 2x10 4 CFU / mL.Item Desenvolvimento de protótipos de testes imunocromatográficos para detecção de anticorpos específicos de leptospiras patogênicas em ratos e cães(Universidade Federal de Goiás, 2018-07-02) Souza, Dienny Rodrigues de; McBride, Alan John Alexander; http://lattes.cnpq.br/2992146566426154; Bührer, Samira; http://lattes.cnpq.br/6715236625981869; Bührer, Samira; Kpnis, André; Scaliante, Rodrigo; Souza, Guilherme Rocha Lino de; Pinto, Emerith Mayra HungriaLeptospirosis is a zoonosis caused by pathogenic spirochetes belonging to the genus Leptospira. The disease may progress to acute renal failure (Weil's Syndrome) and severe pulmonary haemorrhagic syndrome (SPHS), with a mortality rate of > 50%. The highest incidence of the disease occurs among the poorer populations and is associated with the lack of basic sanitation that gets worse during periods of heavy rainfall and flooding. Rodents and dogs are the main sources of transmission of the disease. Therefore, the identification of infected animals is essential towards eliminating reservoir hosts such as dogs and rodents. Therefore, means allowing the identification of leptospira infection in dogs and rodents is essential to permit control actions, decreasing the infection rate in the environment and consequently helping to prevent disease in humans. The objective of this study was to develop prototypes of lateral flow immunochromatographic tests to aid in the diagnosis and control of leptospirosis. The design used in the prototypes was based on a lateral flow immunochromatographic test for direct detection of specific antibodies using a recombinant antigen, known as rLigBrep, attached to the nitrocellulose membrane test line. As an antigen/antibody binding marker, polyvalent antibodies were conjugated to 40 nm colloidal gold, murine anti-mouse immunoglobulin for the murine, anti-canine (Rockland) and anti-canine IgG (Jackson) for canine prototypes. The canine prototypes were tested with 10 positive samples and 10 negative samples previously confirmed by the MAT and ELISA for canine leptospirosis. The staining intensity was recorded on a scale of 0 to 3. Low concentrations of rLigBrep antigen resulted in weak staining intensity. The results of the murine prototype provided the basic specifications to developed the canine prototypes. In the first canine prototype using Rockland conjugate, 5/10 positive samples presented staining intensity of 1. In the canine prototype using Jackson conjugate, 9/10 positive samples presented staining intensity variation of 1 to 3. The results of negative samples 8/10 presented a tenuous staining intensity on the test line, indicating the necessity to reduce the concentration of antigen and/or conjugate on the test. The studied prototypes were not evaluated to determine sensibility and specificity as they didn’t present similar results obtained on ELISA and MAT. Therefore, it has not been defined if the rLigBrep protein can be utilized in the development of a rapid immunochromatography test for the diagnosis of canine leptospirosis and as a marker of infection in rodents.