DEACL - Departamento de Análises Clínicas

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Browsing DEACL - Departamento de Análises Clínicas by Author "Alessio, Glaucia Diniz"

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    Accomplishing the genotype-specific serodiagnosis of single and dual Trypanosoma cruzi infections by flow cytometry Chagas- Flow ATE-IgG2a.
    (2018) Alessio, Glaucia Diniz; Araújo, Fernanda Fortes de; Sales Júnior, Policarpo Ademar; Gomes, Matheus de Souza; Amaral, Laurence Rodrigues do; Xavier, Marcelo Antônio Pascoal; Carvalho, Andréa Teixeira de; Lana, Marta de; Martins Filho, Olindo Assis
    The methods currently available for genotype-specific diagnosis of T. cruzi infection still present relevant limitations, especially to identify mixed infection. In the present investigation, we have evaluated the performance of Chagas-Flow ATE-IgG2a test for early and late differential diagnosis of single and dual genotype-specific T. cruzi infections. Serum samples from Swiss mice at early and late stages of T. cruzi infection were assayed in parallel batches for genotype-specific diagnosis of single (TcI, TcVI or TcII) and dual (TcI+TcVI, TcVI+TcII or TcII+TcI) infections. The intrinsic reactivity to TcI, TcVI and TcII target antigens, including amastigote (AI/AVI/AII), trypomastigote-(TI/TVI/TII) and epimastigote (EI/ EVI/EII), at specific reverse of serum dilutions (500 to 64,000), was employed to provide reliable decision-trees for ªearlyº vs ªlateº, ªsingle vs ªdualº and ªgenotype-specificº serology. The results demonstrated that selective set of attributes ªEII 500/EI 2,000/AII 500º were able to provide high-quality accuracy (81%) to segregate early and late stages of T. cruzi infection. The sets ªTI 2,000/AI 1,000/EII 1,000º and ªTI 8,000/AII 32,000º presented expressive scores to discriminate single from dual T. cruzi infections at early (85%) and late stages (84%), respectively. Moreover, the attributes ªTI 4,000/TVI 500/TII 1,000º, ªTI 16,000/EI 2,000/EII 2,000/AI 500/TVI 500º showed good performance for genotype-specific diagnosis at early stage of single (72%) and dual (80%) T. cruzi infections, respectively. In addition, the attributes ªTI 4,000/AII 1,000/EVI 1,000º, ªTI 64,000/AVI 500/AI 2,000/AII 1,000/EII 4,000º showed moderate performance for genotype-specific diagnosis at late stage of single (69%) and dual (76%) T. cruzi infections, respectively. The sets of decision-trees were assembled to construct a sequential algorithm with expressive accuracy (81%) for serological diagnosis of T. cruzi infection. These findings engender new perspectives for the application of Chagas-Flow ATE-IgG2a method for genotype-specific diagnosis in humans, with relevant contributions for epidemiological surveys as well as clinical and post-therapeutic monitoring of Chagas disease.
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    Innovations in diagnosis and post-therapeutic monitoring of Chagas disease : simultaneous flow cytometric detection of IgG1 antibodies anti-live amastigote, anti-live trypomastigote, and anti-fixed epimastigote forms of Trypanosoma cruzi.
    (2014) Alessio, Glaucia Diniz; Côrtes, Denise Fonseca; Assis, Girley Francisco Machado de; Sales Júnior, Policarpo Ademar; Ferro, Eloisa Amália Vieira; Antonelli, Lis Ribeiro do Valle; Carvalho, Andréa Teixeira de; Martins Filho, Olindo Assis; Lana, Marta de
    This study developed a remarkable methodological innovation (FC-ATE) which enables simultaneous detection of antibodies specific to the three evolutive forms of Trypanosoma cruzi: live amastigote (AMA), live trypomastigote (TRYPO), and fixed epimastigote (EPI) using a differential fluorescence staining as low (AMA), intermediate (TRYPO), and high (EPI). An outstanding performance (100%) was observed in the discrimination of the chagasic (CH) and non-chagasic (NCH) patients. In the applicability of FC-ATE in the diagnosis of Chagas disease, 100% of the CH samples presented positivity in the percentage of positive fluorescent parasites (PPFP) for all the three forms of T. cruzi. Moreover, 94% of the samples of NCH presented negative values of PPFP with AMA and TRYPO, and 88% with EPI. Samples from the NCH group with falsepositive results were those belonging to the leishmaniasis patients. Considering the applicability of this technique in post-therapeuticmonitoring of Chagas disease, 100% of non-treated (NT) and treated non-cured (TNC) samples were positive with the three T. cruzi evolutive forms, while a percentage of 100% fromsamples of the treated cured (TC) patientswere negativewith AMA, 93% with TRYPO and 96% with EPI. The comparison between FC-ATE and two other flow cytometric tests using the same samples of patients NT, TNC and TC showed that the three techniques presented different reactivities, although categorical correlation between the methodologies was observed. Taken together, the results obtained with the novel FC-ATE method have shown an outstanding performance in the diagnosis and post-therapeutic monitoring of Chagas disease.
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    Performance of TcI/TcVI/TcII Chagas-Flow ATE-IgG2a for universal and genotype-specific serodiagnosis of Trypanosoma cruzi infection.
    (2017) Alessio, Glaucia Diniz; Araújo, Fernanda Fortes de; Côrtes, Denise Fonseca; Sales Júnior, Policarpo Ademar; Lima, Daniela Cristina; Gomes, Matheus de Souza; Amaral, Laurence Rodrigues do; Xavier, Marcelo Antônio Pascoal; Carvalho, Andréa Teixeira de; Martins Filho, Olindo Assis; Lana, Marta de
    Distinct Trypanosoma cruzi genotypes have been considered relevant for patient management and therapeutic response of Chagas disease. However, typing strategies for genotype- specific serodiagnosis of Chagas disease are still unavailable and requires standardization for practical application. In this study, an innovative TcI/TcVI/TcII Chagas Flow ATE-IgG2a technique was developed with applicability for universal and genotypespecific diagnosis of T. cruzi infection. For this purpose, the reactivity of serum samples (percentage of positive fluorescent parasites-PPFP) obtained from mice chronically infected with TcI/Colombiana, TcVI/CL or TcII/Y strain as well as non-infected controls were determined using amastigote-AMA, trypomastigote-TRYPO and epimastigote-EPI in parallel batches of TcI, TcVI and TcII target antigens. Data demonstrated that ªα-TcII-TRYPO/ 1:500, cut-off/PPFP = 20%º presented an excellent performance for universal diagnosis of T. cruzi infection (AUC = 1.0, Se and Sp = 100%). The combined set of attributes ªα-TcITRYPO/ 1:4,000, cut-off/PPFP = 50%º, ªα-TcII-AMA/1:1,000, cut-off/PPFP = 40%º and ªα- TcVI-EPI/1:1,000, cut-off/PPFP = 45%º showed good performance to segregate infections with TcI/Colombiana, TcVI/CL or TcII/Y strain. Overall, hosts infected with TcI/Colombiana and TcII/Y strains displayed opposite patterns of reactivity with ªα-TcI TRYPOº and ªα-TcII AMAº. Hosts infected with TcVI/CL strain showed a typical interweaved distribution pattern. The method presented a good performance for genotype-specific diagnosis, with global accuracy of 69% when the population/prototype scenario include TcI, TcVI and TcII infections and 94% when comprise only TcI and TcII infections. This study also proposes a receiver operating reactivity panel, providing a feasible tool to classify serum samples from hosts infected with distinct T. cruzi genotypes, supporting the potential of this method for universal and genotype-specific diagnosis of T. cruzi infection.
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    TcI, TcII and TcVI Trypanosoma cruzi samples from Chagas diseasepatients with distinct clinical forms and critical analysis of in vitro andin vivo behavior, response to treatment and infection evolution inmurine model.
    (2017) Oliveira, Maykon Tavares de; Branquinho, Renata Tupinambá; Alessio, Glaucia Diniz; Mello, Carlos Geraldo Campos de; Paiva, Nívia Carolina Nogueira de; Carneiro, Cláudia Martins; Toledo, Max Jean de Ornelas; Reis, Alexandre Barbosa; Martins Filho, Olindo Assis; Lana, Marta de
    tThe clonal evolution of Trypanosoma cruzi sustains scientifically the hypothesis of association betweenparasite’s genetic, biological behavior and possibly the clinical aspects of Chagas disease in patientsfrom whom they were isolated. This study intended to characterize a range of biological properties ofTcI, TcII and TcVI T. cruzi samples in order to verify the existence of these associations. Several biologicalfeatures were evaluated, including in vitro epimastigote-growth, “Vero”cells infectivity and growth, alongwith in vivo studies of parasitemia, polymorphism of trypomastigotes, cardiac inflammation, fibrosis andresponse to treatment by nifurtimox during the acute and chronic murine infection. The global resultsshowed that the in vitro essays (acellular and cellular cultures) TcII parasites showed higher values for allparameters (growth and infectivity) than TcVI, followed by TcI. In vivo TcII parasites were more virulentand originated from patients with severe disease. Two TcII isolates from patients with severe pathologywere virulent in mice, while the isolate from a patient with the indeterminate form of the disease causedmild infection. The only TcVI sample, which displayed low values in all parameters evaluated, was alsooriginated of an indeterminate case of Chagas disease. Response to nifurtimox was not associated toparasite genetic and biology, as well as to clinical aspects of human disease. Although few number of T.cruzi samples have been analyzed, a discreet correlation between parasite genetics, biological behaviorin vitro and in vivo (murine model) and the clinical form of human disease from whom the samples wereisolated was verified.