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Analysis of the efficiency of a storage system for small amounts of biological material in card, associated with a method of diagnosis and molecular quantification of last generation


Recent epidemic outbreaks demonstrate that adequate and effective interventions in acute care and in the next phases of emerging and reemerging diseases depend on a rapid and reliable detection of the pathogen. There is increasing demand for diagnostic techniques that can determine the infectious pathogen in a sensitive and specific manner, and quantify it accurately in the early stages of the infection. It is also necessary to ensure the safety and easiness of protocols for collection, storage, transport and handling of biological samples. In the present research project, we intend to evaluate the feasibility of a system that integrates collection, storage and transportation of biological samples in cards, together with the identification and quantification of nucleic acid, using digital PCR (dPCR). Initially we will perform digital PCR (dPCR) in DNA samples from HTLV-1 and HTLV-2 seropositive patients, already diagnosed by real-time PCR (RT-qPCR), the current gold standard technique for identifying and quantifying nucleic acids. Comparison of the results obtained by these two techniques will be used to investigate whether the results can be considered similar. In the second stage of the research, we will collect blood samples from HTLV-1 and HTLV-2 seropositive patients, which will be divided into 2 parts: one part will be directly processed for DNA isolation, and the other will be applied in cards, which will be processed for DNA isolation. Then, the DNA samples obtained from both procedures will be used for RT-qPCR and dPCR with specific kits. Our expectation is that dPCR can bring better results than RT-qPCR and the cards can function as a good tool for collection, storage and transport of blood samples without harming the results of PCR techniques. We thus hope to develop a flexible system that will allow our laboratory (DGLab) to offer an efficient molecular diagnostic method (more rapid, sensitive, accurate and with competitive prices), that can meet even the farthest area of Brazil. The potential of the company can be further expanded to include other types of genetic diagnosis of pathogens and other non-infectious diseases. Once reaching our goals, we will have in hand a more practical, secure, democratic, and environmentally friendly (not to use tubes) system for high quality molecular diagnostic. (AU)

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