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PCR-tips for rapid diagnosis of bacterial pathogens

ABSTRACT : 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 dispo...

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Autores principales: Gimenez, Claudia, Sánchez, Mirna L., Valdez, Hugo A., Rodriguez, María E., Grasselli, Mariano
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9343218/
https://www.ncbi.nlm.nih.gov/pubmed/35915170
http://dx.doi.org/10.1007/s00253-022-12069-9
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author Gimenez, Claudia
Sánchez, Mirna L.
Valdez, Hugo A.
Rodriguez, María E.
Grasselli, Mariano
author_facet Gimenez, Claudia
Sánchez, Mirna L.
Valdez, Hugo A.
Rodriguez, María E.
Grasselli, Mariano
author_sort Gimenez, Claudia
collection PubMed
description ABSTRACT : 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. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00253-022-12069-9.
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spelling pubmed-93432182022-08-02 PCR-tips for rapid diagnosis of bacterial pathogens Gimenez, Claudia Sánchez, Mirna L. Valdez, Hugo A. Rodriguez, María E. Grasselli, Mariano Appl Microbiol Biotechnol Methods and Protocols ABSTRACT : 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. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00253-022-12069-9. Springer Berlin Heidelberg 2022-08-02 2022 /pmc/articles/PMC9343218/ /pubmed/35915170 http://dx.doi.org/10.1007/s00253-022-12069-9 Text en © The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Methods and Protocols
Gimenez, Claudia
Sánchez, Mirna L.
Valdez, Hugo A.
Rodriguez, María E.
Grasselli, Mariano
PCR-tips for rapid diagnosis of bacterial pathogens
title PCR-tips for rapid diagnosis of bacterial pathogens
title_full PCR-tips for rapid diagnosis of bacterial pathogens
title_fullStr PCR-tips for rapid diagnosis of bacterial pathogens
title_full_unstemmed PCR-tips for rapid diagnosis of bacterial pathogens
title_short PCR-tips for rapid diagnosis of bacterial pathogens
title_sort pcr-tips for rapid diagnosis of bacterial pathogens
topic Methods and Protocols
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9343218/
https://www.ncbi.nlm.nih.gov/pubmed/35915170
http://dx.doi.org/10.1007/s00253-022-12069-9
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