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Multiplex detection of meningitis pathogens by a vertical flow paper microarray and signal enhancement suitable for low-resource settings: Proof of concept
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Nano Biotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0001-7454-7189
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Nano Biotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab. Department of Global Public Health, Karolinska Institutet, Stockholm, Sweden, Stockholm.ORCID iD: 0000-0003-4923-6965
MSF Epicentre Mbarara Research Centre, Mbarara, Uganda.ORCID iD: 0000-0002-5440-7100
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Nano Biotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0003-4727-6138
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2024 (English)In: Talanta Open, E-ISSN 2666-8319, Vol. 10, article id 100357Article in journal (Refereed) Published
Abstract [en]

Objectives: Meningitis is a medical emergency, and it is crucial to diagnose it accurately and promptly in order to manage patients effectively. It would, therefore, be essential to introduce and have fast, accurate, and user-friendly methods to determine the cause of these infections. This study aimed to demonstrate a potentially cost-effective new approach for detecting meningitis using a paper-based vertical flow microarray, which could be useful in settings with limited resources. Methods: We describe a multiplex paper microarray for detecting Neisseria meningitidis, Haemophilus influenzae, Streptococcus pneumoniae, and Salmonella spp. by the passive vertical flow of PCR-amplified clinical samples. A multibiotinylated amplicon was obtained as a product of PCR in the presence of both a biotinylated primer and biotin-11-dUTP. An enhancement step based on an enzyme-free gold enhancement protocol was also used to facilitate visual detection. Results: This study showed that the vertical flow microarray (previously evaluated for one pathogen) can discriminately detect the amplification results down to the 102 copies of DNA limit for four meningitis pathogens in a multiplexed set-up. The study further demonstrated the ability of this device and setup to detect three of the four pathogens from clinical biosamples. Discussion: This study demonstrated the capacity of a vertical flow microarray device to detect amplification products for four prevalent meningitis pathogens in a multiplex format. The vertical flow microarray demonstrated consistent visualization of the expected gene amplification results; however, indicating limitations in the pre- and amplification steps. This study highlights the potential of this multiplexing method for diagnosing meningitis and other syndromic diseases caused by various pathogens, especially in resource-limited areas.

Place, publisher, year, edition, pages
Elsevier BV , 2024. Vol. 10, article id 100357
Keywords [en]
Global health, Low-resource settings, Multiplex paper microarray, Paediatrics, Passive vertical flow, Point-of-care, Signal enhancement
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:kth:diva-353912DOI: 10.1016/j.talo.2024.100357ISI: 001318148500001Scopus ID: 2-s2.0-85204042669OAI: oai:DiVA.org:kth-353912DiVA, id: diva2:1900987
Note

QC 20241009

Available from: 2024-09-25 Created: 2024-09-25 Last updated: 2025-03-13Bibliographically approved

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Reu, PedroGaudenzi, GiuliaSvedberg, GustavUrrutia Iturritza, MirenGantelius, JesperSvahn Andersson, HeleneRussom, Aman

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Reu, PedroGaudenzi, GiuliaNanjebe, DeborahSvedberg, GustavNyehangane, DanUrrutia Iturritza, MirenMlotshwa, PhuthumaniGantelius, JesperMwanga-Amumpaire, JulietSvahn Andersson, HeleneKumbakumba, EliasRussom, Aman
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