Immunomagnetic separation improves the detection of mycobacteria by paper-based lateral and vertical flow immunochromatographic assays
Ben Aissa Soler, Alejandra (Universitat Autònoma de Barcelona. Departament de Química)
Julián Gómez, Esther (Universitat Autònoma de Barcelona. Departament de Genètica i de Microbiologia)
Zanoni, Maria Valnice (São Paulo State University. Department of Analytical Chemistry)
Pividori, María Isabel (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")

Data:	2021
Resum:	This work addresses a method that combines immunomagnetic separation (IMS) and paperbased nucleic acid immunochromatographic assay for the sensitive detection of Mycolicibacterium fortuitum (basonym Mycobacterium fortuitum) In particular, the preconcentration of the bacteria was achieved by using magnetic particles modified with an antibody specific towards mycobacteria. Following the IMS, the bacteria were lysed, and the genome was amplified by double-tagging PCR, using a set of primers specific for the 16S rRNA gene for Mycobacterium. During the amplification, the amplicons were labeled with biotin and digoxigenin tags. Moreover, a comparative study of paper-based immunochromatographic platforms, relying on vertical and lateral flow and on the use of streptavidin gold nanoparticles as a signal generating system, was also performed. The visual readout was achieved when the gold-modified amplicons were captured by the anti-DIG antibody in the test line. The analytical performance of both methods, nucleic acid vertical flow (NAVF) and nucleic acid lateral flow (NALF), is also discussed. Although NALF showed lower limit of detections (LODs), both NALF and NAVF combined with IMS were able to detect the required LOD in hemodialysis water, becoming two promising and useful techniques for the rapid screening of water supplies in hemodialysis centers, to prevent the exposure of immunosuppressed patients to contaminated sources.
Ajuts:	Agencia Estatal de Investigación PID2019-106625RB-I00 Agència de Gestió d'Ajuts Universitaris i de Recerca 2017/SGR-229
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	Rapid diagnostic tests ; Lateral flow assay ; Vertical flow assay ; Gold nanoparticles ; Mycobacteria ; Double-tagging PCR
Publicat a:	Sensors (Basel, Switzerland), Vol. 21, Issue 18 (September 2021) , art. 5992, ISSN 1424-8220

DOI: 10.3390/s21185992
PMID: 34577201

13 p, 2.6 MB

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Documents de recerca > Documents dels grups de recerca de la UAB > Centres i grups de recerca (producció científica) > Ciències de la salut i biociències > Institut de Biotecnologia i de Biomedicina (IBB)
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