LIVIA DE SOUZA BOTELHO LIMA

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LIM/07 - Laboratório de Gastroenterologia Clínica e Experimental, Hospital das Clínicas, Faculdade de Medicina

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  • article 34 Citação(ões) na Scopus
    Evaluation of real-time PCR assay to detect Schistosoma mansoni infections in a low endemic setting
    (2014) ESPIRITO-SANTO, Maria Cristina Carvalho; ALVARADO-MORA, Monica Viviana; DIAS-NETO, Emmanuel; BOTELHO-LIMA, Livia Souza; MOREIRA, Joao Paulo; AMORIM, Maria; PINTO, Pedro Luiz Silva; HEATH, Ashley R.; CASTILHO, Vera Lucia Pagliusi; GONCALVES, Elenice Messias do Nascimento; LUNA, Expedito Jose de Albuquerque; CARRILHO, Flair Jose; PINHO, Joao Renato Rebello; GRYSCHEK, Ronaldo Cesar Borges
    Background: Schistosomiasis constitutes a major public health problem, and 200 million people are estimated to be infected with schistosomiasis worldwide. In Brazil, schistosomiasis has been reported in 19 states, showing areas of high and medium endemicity and a wide range of areas of low endemicity (ALE). Barra Mansa in Rio de Janeiro state has an estimated prevalence of 1%. ALE represent a new challenge for the helminth control because about 75% of infected individuals are asymptomatic and infections occur with a low parasite load (<100 eggs per gram of feces), causing a decrease in sensitivity of stool parasitological techniques, which are a reference for the laboratory diagnosis of this helminth. The objective of this study was to evaluate the performance of a TaqMan quantitative polymerase chain reaction (qPCR) technique in serum and feces DNA samples using the techniques of Kato-Katz (KK), Hoffman, Pons and Janer (HH) as references, during an epidemiological survey using fecal samples and sera from randomized residents from an ALE. Methods: A cross-sectional study conducted from April to December 2011 using a probabilistic sampling that collected 572 fecal and serum samples. The laboratory diagnostic techniques used were: KK, HH and qPCR ( feces and serum). Results: We obtained the following results using the different diagnostic techniques: KK and HH, 0.9% (n = 5); qPCR-feces, 9.6% (n = 55); and qPCR-serum, 1.4% (n = 8). The qPCR-feces presented the highest positivity, whereas the techniques of HH and KK were the least sensitive to detect infections (0.8%). Compared to HH and KK, qPCR-feces showed a statistically significant difference in positivity (p < 0.05), although with poor agreement. Conclusion: The positivity rate presented by the qPCR approach was far higher than that obtained by parasitological techniques. The lack of adequate surveillance in ALE of schistosomiasis indicates a high possibility of these areas being actually of medium and high endemicity. This study presents a control perspective, pointing to the possibility of using combined laboratory tools in the diagnosis of schistosomiasis in ALE.
  • article 10 Citação(ões) na Scopus
    Detection of Schistosoma mansoni infection by TaqMan (R) Real-Time PCR in a hamster model
    (2014) ESPIRITO-SANTO, Maria Cristina Carvalho; ALVARADO-MORA, Monica Viviana; PINTO, Pedro Luiz Silva; BRITO, Thales de; BOTELHO-LIMA, Livia; HEATH, Ashley Richard; AMORIM, Maria Galli; DIAS-NETO, Emmanuel; CHIEFFI, Pedro Paulo; PINHO, Joao Renato Rebello; CARRILHO, Flair Jose; LUNA, Expedito Jose Albuquerque; GRYSCHEK, Ronaldo Cesar Borges
    An experimental study in hamsters was performed to evaluate the capability for detecting Schistosoma mansoni DNA in serum and fecal samples during the pre and post-egg-laying periods of infection using TaqMan (R) Real-Time PCR system (qPCR), was compared with the circumoval precipitin test (COPT) and the Kato-Katz technique, especially among individuals with low parasitic burden. Twenty-four hamsters were infected with cercariae. Three hamsters were sacrificed per week under anesthesia, from 7 days post infection (DPI) up to 56 DPI. A serum sample and a pool of feces were collected from each hamster. The presence of S. mansoni eggs in fecal samples was evaluated by Kato-Katz method and in the hamsters gut-by histopathology. Detection of S. mansoni DNA was performed using qPCR and S. mansoni antibody using COPT. The first detection of eggs in feces by Kato-Katz method and S. mansoni DNA in feces by qPCR occurred 49 DPI. Nevertheless, S. mansoni DNA was detected in serum samples from 14 up to 56 DPI. COPT was positive at 35 DPI. The results not only confirm the reliability of S. mansoni DNA detection by qPCR, but also demonstrate that serum is a trustworthy source of DNA in the pre patent infection period.