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A Trypanosoma cruzi Genome Tandem Repetitive Satellite DNA Sequence as a Molecular Marker for a LAMP Assay for Diagnosing Chagas' Disease
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Autores
Ordóñez, Diego
Fernández-Soto, Pedro
Fernández-Martín, Ana M
Crego-Vicente, Beatriz
Febrer-Sendra, Begoña
Diego, Juan García-Bernalt
Vicente, Belén
López-Abán, Julio
Belhassen-García, Moncef
Muro, Antonio
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Fecha
2020
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Hindawi Limited
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Abstract
Chagas' disease is a neglected tropical disease caused by Trypanosoma cruzi which is endemic throughout Latin America and is spread by worldwide migration. Diagnosis is currently limited to serological and molecular techniques having variations regarding their sensitivity and specificity. This work was aimed at developing a new sensitive, applicable, and cost-effective molecular diagnosis technique for loop-mediated isothermal amplification-based detection of T. cruzi (Tc-LAMP). The results led to determining a highly homologous satellite repeat region (231 bp) among parasite strains as a molecular marker for diagnosing the disease. Tc-LAMP was performed correctly for detecting parasite DNA (5 fg for the CL Brener strain and 50 fg for the DM28, TcVI, and TcI strains). Assay results proved negative for DNA from 16 helminth species and 7 protozoa, including Leishmania spp. Tc-LAMP based on the highly repeated T. cruzi satellite region is thus proposed as an important alternative for diagnosing T. cruzi infection, overcoming other methods' limitations such as their analytic capability, speed, and requiring specialized equipment or highly trained personnel. Tc-LAMP could be easily adapted for point-of-care testing in areas having limited resources. © 2020 Diego Ordóñez et al.
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Keywords
Calcium binding protein , Ribosome rna , Satellite dna , Article , Chagas disease , Colorimetry , Controlled study , Cost effectiveness analysis , Cross reaction , Dna contamination , Dna template , Human , In vitro study , Limit of detection , Loop mediated isothermal amplification , Molecular diagnosis , Nonhuman , Nucleotide sequence , Point of care testing , Real time polymerase chain reaction , Trypanosoma cruzi
