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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, Skolan för kemi, bioteknologi och hälsa (CBH), Proteinvetenskap, Nanobioteknologi. KTH, Centra, Science for Life Laboratory, SciLifeLab.ORCID-id: 0000-0001-7454-7189
KTH, Skolan för kemi, bioteknologi och hälsa (CBH), Proteinvetenskap, Nanobioteknologi. KTH, Centra, 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, Skolan för kemi, bioteknologi och hälsa (CBH), Proteinvetenskap, Nanobioteknologi. KTH, Centra, Science for Life Laboratory, SciLifeLab.ORCID-id: 0000-0003-4727-6138
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2024 (Engelska)Ingår i: Talanta Open, E-ISSN 2666-8319, Vol. 10, artikel-id 100357Artikel i tidskrift (Refereegranskat) 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.

Ort, förlag, år, upplaga, sidor
Elsevier BV , 2024. Vol. 10, artikel-id 100357
Nyckelord [en]
Global health, Low-resource settings, Multiplex paper microarray, Paediatrics, Passive vertical flow, Point-of-care, Signal enhancement
Nationell ämneskategori
Medicin och hälsovetenskap
Identifikatorer
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
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QC 20241009

Tillgänglig från: 2024-09-25 Skapad: 2024-09-25 Senast uppdaterad: 2025-03-13Bibliografiskt granskad

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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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NanobioteknologiScience for Life Laboratory, SciLifeLab
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