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High-Throughput 2018, 7(4), 32; https://doi.org/10.3390/ht7040032

Development and Assessment of a Diagnostic DNA Oligonucleotide Microarray for Detection and Typing of Meningitis-Associated Bacterial Species

1
Public Health England, Porton Down, Salisbury SP4 0JG, UK
2
Department of Clinical Microbiology, Royal Cornwall Hospitals NHS Trust, Penventinnie Lane, Treliske, Truro, Cornwall TR1 3LQ, UK
3
Public Health England Laboratory Southampton, Southampton General Hospital, Tremona Road, Southampton SO16 6YD, UK
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Received: 13 August 2018 / Revised: 11 September 2018 / Accepted: 21 September 2018 / Published: 16 October 2018
(This article belongs to the Special Issue Applications of Microarrays in Diagnostics)
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Abstract

Meningitis is commonly caused by infection with a variety of bacterial or viral pathogens. Acute bacterial meningitis (ABM) can cause severe disease, which can progress rapidly to a critical life-threatening condition. Rapid diagnosis of ABM is critical, as this is most commonly associated with severe sequelae with associated high mortality and morbidity rates compared to viral meningitis, which is less severe and self-limiting. We have designed a microarray for detection and diagnosis of ABM. This has been validated using randomly amplified DNA targets (RADT), comparing buffers with or without formamide, in glass slide format or on the Alere ArrayTubeTM (Alere Technologies GmbH) microarray platform. Pathogen-specific signals were observed using purified bacterial nucleic acids and to a lesser extent using patient cerebral spinal fluid (CSF) samples, with some technical issues observed using RADT and glass slides. Repurposing the array onto the Alere ArrayTubeTM platform and using a targeted amplification system increased specific and reduced nonspecific hybridization signals using both pathogen nucleic and patient CSF DNA targets, better revealing pathogen-specific signals although sensitivity was still reduced in the latter. This diagnostic microarray is useful as a laboratory diagnostic tool for species and strain designation for ABM, rather than for primary diagnosis. View Full-Text
Keywords: meningitis; microarray; bacterial; diagnostic; assay; ArrayTube; diagnosis; infection meningitis; microarray; bacterial; diagnostic; assay; ArrayTube; diagnosis; infection
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Bannister, S.A.; Kidd, S.P.; Kirby, E.; Shah, S.; Thomas, A.; Vipond, R.; Elmore, M.J.; Telfer Brunton, A.; Marsh, P.; Green, S.; Silman, N.J.; Kempsell, K.E. Development and Assessment of a Diagnostic DNA Oligonucleotide Microarray for Detection and Typing of Meningitis-Associated Bacterial Species. High-Throughput 2018, 7, 32.

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