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|Title: ||Loop-Mediated Isothermal Amplification for Laboratory Confirmation of Buruli Ulcer Disease—Towards a Point-of-Care Test|
|Authors: ||Beissner, Marcus|
Phillips, Richard Odame
Sarfo, Fred Stephen
|Issue Date: ||2015|
|Publisher: ||Plos Neglected Tropical Diseases|
|Citation: ||Beissner M, Phillips RO, Battke F, Bauer M, Badziklou K, Sarfo FS, et al. (2015) Loop-Mediated Isothermal Amplification for Laboratory Confirmation of Buruli Ulcer Disease—Towards a Point-of-Care Test. PLoS Negl Trop Dis 9(11): e0004219. doi:10.1371/journal.pntd.0004219|
As the major burden of Buruli ulcer disease (BUD) occurs in remote rural areas, development
of point-of-care (POC) tests is considered a research priority to bring diagnostic services
closer to the patients. Loop-mediated isothermal amplification (LAMP), a simple,
robust and cost-effective technology, has been selected as a promising POC test candidate.
Three BUD-specific LAMP assays are available to date, but various technical challenges
still hamper decentralized application. To overcome the requirement of cold-chains
for transport and storage of reagents, the aim of this study was to establish a dry-reagentbased
LAMP assay (DRB-LAMP) employing lyophilized reagents.
Following the design of an IS2404 based conventional LAMP (cLAMP) assay suitable to
apply lyophilized reagents, a lyophylization protocol for the DRB-LAMP format was developed.
Clinical performance of cLAMP was validated through testing of 140 clinical samples
from 91 suspected BUD cases by routine assays, i.e. IS2404 dry-reagent-based (DRB) PCR,
conventional IS2404 PCR (cPCR), IS2404 qPCR, compared to cLAMP.Whereas qPCR rendered
an additional 10%of confirmed cases and samples respectively, case confirmation
and positivity rates of DRB-PCR or cPCR (64.84%and 56.43%; 100%concordant results in
both assays) and cLAMP (62.64%and 52.86%) were comparable and there was no significant
difference between the sensitivity of the assays (DRB PCR and cPCR, 86.76%; cLAMP,83.82%). Likewise, sensitivity of cLAMP (95.83%) and DRB-LAMP (91.67%) were comparable
as determined on a set of 24 samples tested positive in all routine assays.
Both LAMP formats constitute equivalent alternatives to conventional PCR techniques. Provided
the envisaged availability of field friendly DNA extraction formats, both assays are
suitable for decentralized laboratory confirmation of BUD, whereby DRB-LAMP scores with
the additional advantage of not requiring cold-chains. As validation of the assays was conducted
in a third-level laboratory environment, field based evaluation trials are necessary to
determine the clinical performance at peripheral health care level.|
|Description: ||An article published by Beissner M, Phillips RO, Battke F, Bauer M,
Badziklou K, Sarfo FS, et al. (2015) Loop-Mediated
Isothermal Amplification for Laboratory Confirmation
of Buruli Ulcer Disease—Towards a Point-of-Care
Test. PLoS Negl Trop Dis 9(11): e0004219.
|Appears in Collections:||College of Health Sciences|
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