Publication: Development of a prototype lateral flow immunoassay (LFI) for the rapid diagnosis of melioidosis.
Accepted Date
2014-01-18
Issued Date
2014-03-20
Resource Type
Language
eng
ISSN
1935-2735 (electronic)
1935-2727 (printed)
1935-2727 (printed)
Rights
Mahidol University
Rights Holder(s)
PubMed Central
Bibliographic Citation
Houghton RL, Reed DE, Hubbard MA, Dillon MJ, Chen H, Currie BJ. et al. Development of a prototype lateral flow immunoassay (LFI) for the rapid diagnosis of melioidosis. PLoS Negl Trop Dis. 2014 Mar 20;8(3):e2727.
Suggested Citation
Houghton, Raymond L., Reed, Dana E., Hubbard, Mark A., Dillon, Michael J., Chen, Hongjing, Currie, Bart J., Mayo, Mark, Sarovich, Derek S., Theobald, Vanessa, Direk Limmathurotsakul, ดิเรก ลิ้มมธุรสกุล, Gumphol Wongsuvan, Narisara Chantratita, นริศรา จันทราทิตย์, Peacock, Sharon J., Hoffmaster, Alex R, Duval, Brea, Brett, Paul J., Burtnick, Mary N., AuCoin, David P. Development of a prototype lateral flow immunoassay (LFI) for the rapid diagnosis of melioidosis.. Houghton RL, Reed DE, Hubbard MA, Dillon MJ, Chen H, Currie BJ. et al. Development of a prototype lateral flow immunoassay (LFI) for the rapid diagnosis of melioidosis. PLoS Negl Trop Dis. 2014 Mar 20;8(3):e2727.. doi:10.1371/journal.pntd.0002727 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/825
Research Projects
Organizational Units
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Title
Development of a prototype lateral flow immunoassay (LFI) for the rapid diagnosis of melioidosis.
Author(s)
Houghton, Raymond L.
Reed, Dana E.
Hubbard, Mark A.
Dillon, Michael J.
Chen, Hongjing
Currie, Bart J.
Mayo, Mark
Sarovich, Derek S.
Theobald, Vanessa
Direk Limmathurotsakul
ดิเรก ลิ้มมธุรสกุล
Gumphol Wongsuvan
Narisara Chantratita
นริศรา จันทราทิตย์
Peacock, Sharon J.
Hoffmaster, Alex R
Duval, Brea
Brett, Paul J.
Burtnick, Mary N.
AuCoin, David P.
Reed, Dana E.
Hubbard, Mark A.
Dillon, Michael J.
Chen, Hongjing
Currie, Bart J.
Mayo, Mark
Sarovich, Derek S.
Theobald, Vanessa
Direk Limmathurotsakul
ดิเรก ลิ้มมธุรสกุล
Gumphol Wongsuvan
Narisara Chantratita
นริศรา จันทราทิตย์
Peacock, Sharon J.
Hoffmaster, Alex R
Duval, Brea
Brett, Paul J.
Burtnick, Mary N.
AuCoin, David P.
Corresponding Author(s)
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.
