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J Mol Diagn ; 21(5): 839-851, 2019 09.
Artículo en Inglés | MEDLINE | ID: mdl-31173930

RESUMEN

Although molecular diagnostics is well established in clinical laboratories, its full potential has not been extended to field settings. Typically, diagnostic real-time quantitative PCR (qPCR) reagents require temperature-controlled transportation and storage. Furthermore, thermocyclers are bulky and fragile, requiring good infrastructure for optimal operation. These major hurdles strongly limit use of molecular-based tests in low-resource scenarios. Herein, Trypanosoma cruzi or Plasmodium spp. DNA were detected with qPCR using commercial equipment (ABI7500 instrument) and a prototype platform comprising a portable device and a silicon chip, named Q3-Plus. In addition, a ready-to-use reaction format, where all qPCR reagents are stored on plate or on chip, was compared with the traditional freezer-stored format. No significant differences were observed in detecting T. cruzi or Plasmodium spp. DNA between thermocyclers, as well as between reagents' formats, for storage periods of up to 28 days (at 2°C to 8°C or 21°C to 23°C, respectively). When challenged with patients' samples, the Q3-Plus system performed as efficiently as the standard equipment for Plasmodium spp. DNA detection, showing it to be a valuable solution to malaria point-of-care diagnostics. Detection of T. cruzi DNA in chronic patients' samples using the Q3-Plus system yielded approximately 50% efficiency relative to the ABI7500. These results are essential to support future endeavors to bring molecular diagnostics to the point of care, where most needed.


Asunto(s)
Enfermedad de Chagas/diagnóstico , ADN Protozoario/análisis , Pruebas Diagnósticas de Rutina/instrumentación , Malaria Falciparum/diagnóstico , Plasmodium falciparum/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Trypanosoma cruzi/genética , Enfermedad de Chagas/parasitología , ADN Protozoario/sangre , ADN Protozoario/genética , Pruebas Diagnósticas de Rutina/métodos , Pruebas Diagnósticas de Rutina/normas , Humanos , Malaria Falciparum/parasitología , Plasmodium falciparum/aislamiento & purificación , Trypanosoma cruzi/aislamiento & purificación
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