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• W233494876 abstract "The rate of infection by Trypanosoma cruzi in triatomines is an important parameter forassessing the risk of vectorial transmission of Chagas disease in localities where domestic andperidomestic infestation by these insects are found. Infection with T. cruzi in triatomines is usuallyevaluated by examining, by light microscopy, suspensions of intestinal contents of fresh insects,seeking in them the presence of flagellate trypanosomatids. This approach has disadvantages as theneed to transport the live insects to the laboratory, and not allow the identification of thetrypanosomatid species present in the triatomine feces. Molecular biology techniques based onpolymerase chain reaction (PCR) have therefore been developed to identify T. cruzi in biologicalsamples, such as the intestinal contents of triatomines. The aim of this study was to evaluate theapplicability and sensitivity of the technique of molecular diagnosis of natural infection by T. cruzi,based on the use of filter paper in triatomines captured in the Ceara state. Additionally, we sought toevaluate this diagnostic technique in samples of intestinal content of triatomines cryopreserved onfilter paper for 11 years. We studied the intestinal contents of 68 specimens of insects captured in theyear 2000 (Rhodnius nasutus, n = 40; T. pseudomaculata, n = 25 and T. brasiliensis, n = 3), and fecesfrom 51 specimens collected in 2010 (T. pseudomaculata, n = 25; T. brasiliensis, n = 36). Theintestinal content obtained by abdominal compression of 119 insects were analyzed by directexamination through optical microscopy. The feces were diluted on microscope slides in bufferedsaline (pH = 7.0), covered with coverslips and examined under optical microscope with 400 timesmagnification for direct observation of the trypanosomes presence. Aliquots of fecal suspensions wereseparated and applied to strips of filter paper (FTA Card, GE Healthcare, Buckinghamshire, UK),which were stored individually in 2 mL tubes and stored at -20 ° C for use in PCR , aimed at detectionof kDNA of T. cruzi. The primers used were 5'-AAATAATGTACGGG S35 (T / G)GAGATGCATGA-3 'and 5'-GGTTGCATTGGGTTGGTGTAATATA S36-3', which amplify afragment of 330 base pairs containing the hypervariable regions of the k-DNA minicircles. Thecomparative calculation of the sensitivity of diagnostic techniques (optical microscopy and PCR) wasperformed as follows: sensitivity = number of technical tests positive by this technique / number ofpositive tests in any of the two techniques. Regarding the analysis of 68 cryopreserved paper intestinalcontents FTA Card, 58 were positive by light microscopy. Of these, 56 were positive by PCR. Wasobtained in this way, a comparative sensitivity of 96.5% for PCR and 100% for lightmicroscopy. Analyzing the newly obtained 51 fecal samples of insects, we obtained a positivity bylight microscopy, four samples and PCR of 14 samples, yielding a sensitivity of 100% for PCR and28.6% for optical microscopy. The agreement between the techniques was 66/68 (97%) for thecryopreserved samples and 41/51 (80.4%) for the newly obtained samples of insects. Analyzing thesamples together, we achieved an agreement of 107/119 (89.9%) between the techniques. The studyconcludes that the technique of PCR for diagnosis in samples of intestinal content of triatominesapplied to FTA paper Card is applicable for estimating natural infection rates of these insects byT. cruzi, is able to assist the characterization of eco-epidemiological profiles and the detection of theparasite in the wild environment, peridomestic and domestic, also contributing to the assessment ofrisk of transmission of Chagas disease in localities where there is infestation by vectors." @default.
• W233494876 created "2016-06-24" @default.
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• W233494876 date "2012-01-01" @default.
• W233494876 modified "2023-09-23" @default.
• W233494876 title "Diagnóstico molecular da infecção natural por Trypanosoma cruzi em triatomíneos nativos de área endêmica para doença de Chagas no estado do Ceará" @default.
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