A colorimetric strategy based on dynamic chemistry for direct detection of Trypanosomatid species
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AuthorTabraue Chávez, Mavys; Luque González, Angélica; Marín Romero, Antonio; Sánchez Martín, Rosario María; Escobedo Araque, Pablo; Pernagallo, Salvatore; Díaz Mochón, Juan José
Tabraue-Chávez, M., Luque-González, M. A., Marín-Romero, A., Sánchez-Martín, R. M., Escobedo-Araque, P., Pernagallo, S., & Díaz-Mochón, J. J. (2019). A colorimetric strategy based on dynamic chemistry for direct detection of Trypanosomatid species. Scientific reports, 9(1), 1-13.
SponsorshipThis research work has received funding from Junta de Andalucía, Consejería de Economía e Innovación (project number 2012-BIO1778), the Spanish Ministerio de Economía y Competitividad (Grants CTQ2012-34778, BIO2016-80519-R, FPI Grant BES-2013- 063020). This research was partially supported by the 7th European Community Framework Program (FP7-PEOPLE-2012-CIG-Project Number 322276).
Leishmaniasis and Chagas disease are endemic in many countries, and re-emerging in the developed countries. A rapid and accurate diagnosis is important for early treatment for reducing the duration of infection as well as for preventing further potential health complications. In this work, we have developed a novel colorimetric molecular assay that integrates nucleic acid analysis by dynamic chemistry (ChemNAT) with reverse dot-blot hybridization in an array format for a rapid and easy discrimination of Leishmania major and Trypanosoma cruzi. The assay consists of a singleplex PCR step that amplifies a highly homologous DNA sequence which encodes for the RNA component of the large ribosome subunit. The amplicons of the two different parasites differ between them by single nucleotide variations, known as “Single Nucleotide Fingerprint” (SNF) markers. The SNF markers can be easily identified by naked eye using a novel micro Spin-Tube device "Spin-Tube", as each of them creates a specific spot pattern. Moreover, the direct use of ribosomal RNA without requiring the PCR pre-amplification step is also feasible, further increasing the simplicity of the assay. The molecular assay delivers sensitivity capable of identifying up to 8.7 copies per μL with single mismatch specificity. The Spin-Tube thus represents an innovative solution providing benefits in terms of time, cost, and simplicity, all of which are crucial for the diagnosis of infectious disease in developing countries.