Development of a Prototype Lateral Flow Immunoassay (LFI) for the Rapid Diagnosis of Melioidosis

R L Houghton; D E Reed; M A Hubbard; M J Dillon; H Chen; B J Currie; M Mayo; Derek S Sarovich; V Theobald; D Limmathurotsakul; G Wongsuvan; N Chantratita; S J Peacock; A R Hoffmaster; B Duval; P J Brett; M N Burtnick; David P AuCoin

doi:10.1371/journal.pntd.0002727

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Development of a Prototype Lateral Flow Immunoassay (LFI) for the Rapid Diagnosis of Melioidosis

Journal article

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Development of a Prototype Lateral Flow Immunoassay (LFI) for the Rapid Diagnosis of Melioidosis

R L Houghton, D E Reed, M A Hubbard, M J Dillon, H Chen, B J Currie, M Mayo, Derek S Sarovich, V Theobald, D Limmathurotsakul, …

PLoS Neglected Tropical Diseases, Vol.8(3), e2727

2014

DOI: https://doi.org/10.1371/journal.pntd.0002727

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Abstract

Burkholderia pseudomallei is a soil-dwelling bacterium and the causative agent of melioidosis. Isolation of B. pseudomallei from clinical samples is the "gold standard" for the diagnosis of melioidosis; results can take 3-7 days to produce. Alternatively, antibody-based tests have low specificity due to a high percentage of seropositive individuals in endemic areas. There is a clear need to develop a rapid point-of-care antigen detection assay for the diagnosis of melioidosis. Previously, we employed In vivo Microbial Antigen Discovery (InMAD) to identify potential B. pseudomallei diagnostic biomarkers. The B. pseudomallei capsular polysaccharide (CPS) and numerous protein antigens were identified as potential candidates. Here, we describe the development of a diagnostic immunoassay based on the detection of CPS. Following production of a CPS-specific monoclonal antibody (mAb), an antigen-capture immunoassay was developed to determine the concentration of CPS within a panel of melioidosis patient serum and urine samples. The same mAb was used to produce a prototype Active Melioidosis Detect Lateral Flow Immunoassay (AMD LFI); the limit of detection of the LFI for CPS is comparable to the antigen-capture immunoassay (~0.2 ng/ml). The analytical reactivity (inclusivity) of the AMD LFI was 98.7% (76/77) when tested against a large panel of B. pseudomallei isolates. Analytical specificity (cross-reactivity) testing determined that 97.2% of B. pseudomallei near neighbor species (35/36) were not reactive. The non-reactive B. pseudomallei strain and the reactive near neighbor strain can be explained through genetic sequence analysis. Importantly, we show the AMD LFI is capable of detecting CPS in a variety of patient samples. The LFI is currently being evaluated in Thailand and Australia; the focus is to optimize and validate testing procedures on melioidosis patient samples prior to initiation of a large, multisite pre-clinical evaluation.

Details

Title: Development of a Prototype Lateral Flow Immunoassay (LFI) for the Rapid Diagnosis of Melioidosis
Authors: R L Houghton (Author) - InBios International Inc, United States
D E Reed (Author) - University of Nevada School of Medicine, United States
M A Hubbard (Author) - University of Nevada School of Medicine, United States
M J Dillon (Author) - University of Nevada School of Medicine, United States
H Chen (Author) - InBios International Inc, United States
B J Currie (Author) - Royal Darwin Hospital
M Mayo (Author) - Royal Darwin Hospital
Derek S Sarovich (Author) - Royal Darwin Hospital
V Theobald (Author) - Royal Darwin Hospital
D Limmathurotsakul (Author) - Mahidol University, Thailand
G Wongsuvan (Author) - Ubon Ratchathani, Thailand
N Chantratita (Author) - Mahidol University, Thailand
S J Peacock (Author) - Mahidol University, Thailand
A R Hoffmaster (Author) - Centers for Disease Control and Prevention, United States
B Duval (Author) - InBios International Inc, United States
P J Brett (Author) - Centers for Disease Control and Prevention, United States
M N Burtnick (Author) - University of South Alabama, United States
David P AuCoin (Author) - University of Nevada School of Medicine, United States
Publication details: PLoS Neglected Tropical Diseases, Vol.8(3), e2727
Publisher: Public Library of Science
Date published: 2014
DOI: 10.1371/journal.pntd.0002727
ISSN: 1935-2727
Copyright note: Copyright © 2014 The Authors. This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.
Organisation Unit: University of the Sunshine Coast, Queensland; Centre for Bioinnovation
Language: English
Record Identifier: 99451166002621
Output Type: Journal article

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