Adenoviral detection by recombinase polymerase amplification and vertical flow paper microarray.
Anal Bioanal Chem
; 411(4): 813-822, 2019 Feb.
Article
en En
| MEDLINE
| ID: mdl-30498984
ABSTRACT
Respiratory viral infections often mimic the symptoms of infections caused by bacteria; however, restricted and targeted administration of antibiotics is needed to combat growing antimicrobial resistance. This is particularly relevant in low-income settings. In this work, we describe the use of isothermal amplification of viral DNA at 37 °C coupled to a paper-based vertical flow microarray (VFM) setup that utilizes a colorimetric detection of amplicons using functionalized gold nanoparticles. Two oligonucleotide probes, one in-house designed and one known adenoviral probe were tested and validated for microarray detection down to 50 nM using a synthetic target template. Furthermore, primers were shown to function in a recombinase polymerase amplification reaction using both synthetic template and viral DNA. As a proof-of-concept, we demonstrate adenoviral detection with four different adenoviral species associated with respiratory infections using the paper-based VFM format. The presented assay was validated with selected adenoviral species using the in-house probe, enabling detection at 1 ng of starting material with intra- and inter-assay %CV of ≤ 9% and ≤ 13%. Finally, we validate our overall method using clinical samples. Based on the results, the combination of recombinase polymerase amplification, paper microarray analysis, and nanoparticle-based colorimetric detection could thus be a useful strategy towards rapid and affordable multiplexed viral diagnostics.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
ADN Viral
/
Adenoviridae
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Técnicas de Amplificación de Ácido Nucleico
/
Recombinasas
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ADN Polimerasa Dirigida por ADN
Tipo de estudio:
Diagnostic_studies
Límite:
Humans
Idioma:
En
Revista:
Anal Bioanal Chem
Año:
2019
Tipo del documento:
Article
País de afiliación:
Suecia