PCR-tips for rapid diagnosis of bacterial pathogens.

Autor: Gimenez C; Departamento de Ciencia y Tecnología, Laboratorio de Materiales Biotecnológicos (LaMaBio), Universidad Nacional de Quilmes, Bernal, Buenos Aires, Argentina.; GBEyB, Multidisciplinary Institute of Cell Biology (IMBICE, Dependent of the Argentine Research Council (CONICET), Scientific Research Commission, Province of Buenos Aires (CIC‑PBA) and UNLP), La Plata, Argentina., Sánchez ML; Departamento de Ciencia y Tecnología, Laboratorio de Materiales Biotecnológicos (LaMaBio), Universidad Nacional de Quilmes, Bernal, Buenos Aires, Argentina.; GBEyB, Multidisciplinary Institute of Cell Biology (IMBICE, Dependent of the Argentine Research Council (CONICET), Scientific Research Commission, Province of Buenos Aires (CIC‑PBA) and UNLP), La Plata, Argentina., Valdez HA; Laboratorio de Microbiología Celular e Inmunomecanismos, CINDEFI | Centro de Investigación y Desarrollo en Fermentaciones Industriales Facultad de Ciencias Exactas (UNLP), La Plata, Argentina., Rodriguez ME; Laboratorio de Microbiología Celular e Inmunomecanismos, CINDEFI | Centro de Investigación y Desarrollo en Fermentaciones Industriales Facultad de Ciencias Exactas (UNLP), La Plata, Argentina., Grasselli M; Departamento de Ciencia y Tecnología, Laboratorio de Materiales Biotecnológicos (LaMaBio), Universidad Nacional de Quilmes, Bernal, Buenos Aires, Argentina. mariano.grasselli@unq.edu.ar.; GBEyB, Multidisciplinary Institute of Cell Biology (IMBICE, Dependent of the Argentine Research Council (CONICET), Scientific Research Commission, Province of Buenos Aires (CIC‑PBA) and UNLP), La Plata, Argentina. mariano.grasselli@unq.edu.ar.
Jazyk: angličtina
Zdroj: Applied microbiology and biotechnology [Appl Microbiol Biotechnol] 2022 Sep; Vol. 106 (17), pp. 5729-5739. Date of Electronic Publication: 2022 Aug 02.
DOI: 10.1007/s00253-022-12069-9
Abstrakt: Micropipette tips are currently among the most used disposable devices in bioresearch and development laboratories. Their main application is the fractionation of solutions. New functionalities have recently been added to this device, widening their applications. This paper analyzed disposable micropipette tips as reagent holders of PCR reagents. PCR has become a prevalent and often indispensable technique in biological laboratories for various applications, such as the detection of coronavirus and other infectious diseases. A functional micropipette tip was implemented to simplify PCR analysis and reduce the contamination chances of deoxynucleotides and specific primers. This disposable device is prepared by tip coating processes of reagents, using polyvinyl alcohol polymer and additives. The coated layer is optimized to load and release PCR reagents efficiently. As a proof of concept, we show that the detection of Bordetella pertussis, the etiological agent of whooping cough whose diagnostic relies on PCR, can be quickly done using practical-functional tips. This device is an excellent example of testing the functionality and contribution of molecular diagnostic PCR tips. KEY POINTS: • Functional micropipette tips are prepared by coating with dNTPs and primers. • Functional tips are used to replace dNTPs and primers in the PCR master mix. • PCR diagnostic of Bordetella pertussis is performed using functional tips.
(© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)
Databáze: MEDLINE
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