Field implementation of a 3D printer based DNA extraction method coupled to LAMP for congenital Chagas disease diagnosis

• Autores: Wehrendt, Diana Patricia; Wong, Season; Rojas Panozo, Lizeth; Rivera Nina, Silvia; Pinto, Lilian; Abril, Marcelo; Lozano, Daniel; Picado, albert; Gascón, Joaquín; Torrico, Faustino; Alonso Padilla, Julio; Schijman, Alejandro Gabriel
• Año de publicación: 2019
• Idioma: inglés
• Tipo de recurso: documento de conferencia
• Materia: Trypanosoma cruzi; Chagas disease; Printrlab; LAMP
• Institución: Consejo Nacional de Investigaciones Científicas y Técnicas

Congenital Chagas disease entails the transmission of Trypanosoma cruzi infection from a mother to her child. With currently available chemotherapies, the cure rate for infected children is almost 100 % if administered early upon infection. It is therefore of great relevance to diagnose newborns on time. However, the algorithm to detect congenital T. cruzi infection involves the performance of microhematocrite or micromethod at delivery or during the first months of life and a confirmatory serology at 10 months of age. In highly endemic areas where people live far away from reference centers, many infants never go back to confirm the diagnosis and receive treatment if infected. The challenge is then to implement sensitive and rapid diagnostic techniques that can be performed in minimally equipped laboratories. At present there is a prototype loop isothermal amplification molecular test available (T. cruzi-LAMP kit, Eiken, Japan), with similar sensitivity to that of real time PCR (qPCR), but easier to use. Nonetheless, highly purified DNA is needed and obtaining it is time consuming and requires equipment unavailable in endemic regions. Thus, our aim was to couple the T. cruzi-LAMP kit to a recently developed DNA extraction device based on a low cost 3D printer (named PrintrLab), and to test its use in a hospital located in the “Gran Chaco”, a highly endemic region for Chagas disease. The PrintrLab was programmed to purify DNA from whole blood-EDTA samples and to provide the incubation step for the T. cruzi-LAMP reaction. The process took about 2.5 hours to yield a result, while manual DNA extraction and subsequent qPCR normally take more than 6. Performance of the “PrintrLab-LAMP” duo was tested with blood-EDTA samples artificially contaminated with 0, 1, 2, 5, 10 and 100 parasites eq/mL and a sensitivity around 2 pararasites eq/mL was achieved. Finally, 70 clinical samples from infants born to seropositive mothers were evaluated and all the micromethod positive ones, 6 samples in total, were detected by the "PrintrLab-LAMP" approach. In conclusion, the “PrintrLabLAMP” device showed a good sensitivity, the protocol was faster than other molecular techniques and it could be successfully used in a minimally equipped laboratory.
Fil: Wehrendt, Diana Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Wong, Season. Ai Biosciences Inc; Estados Unidos
Fil: Rojas Panozo, Lizeth. Ceades; Bolivia
Fil: Rivera Nina, Silvia. Ceades; Bolivia
Fil: Pinto, Lilian. Fundación Mundo Sano; Argentina
Fil: Abril, Marcelo. Fundación Mundo Sano; Argentina
Fil: Lozano, Daniel. Ceades; Bolivia
Fil: Picado, albert. Foundation For Innovative Diagnostics; Suiza
Fil: Gascón, Joaquín. Instituto de Salud Global de Barcelona; España
Fil: Torrico, Faustino. Ceades; Bolivia
Fil: Alonso Padilla, Julio. Instituto de Salud Global de Barcelona; España
Fil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas y VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Mar del Plata
Argentina
Sociedad Argentina de Investigación Clínica
Asociación Argentina de Farmacología Experimental
Sociedad Argentina de Biología
Sociedad Argentina de Protozoología
Asociación Argentina de Nanomedicinas
Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio