Loop-mediated isothermal amplification for laboratory confirmation of Buruli ulcer disease: Towards a point-of-care test.

Author Summary:

Buruli ulcer disease (BUD) mainly occurs in remote rural areas of Sub-Saharan Africa, affects skin and soft tissue, and may lead to severe disabilities. Therefore, early diagnosis and treatment with antimycobacterial therapy are essential whereby the WHO recommends laboratory confirmation of 70% of the cases. As the current diagnostic gold standard (polymerase chain reaction [PCR]) is restricted to third-level laboratories, development of confirmatory point-of-care (POC) tests for BUD applicable at primary health care level has become a research priority to bring diagnosis closer to where the patients are. Loop-mediated isothermal amplification (LAMP) has been selected by the WHO as one of the promising candidate technologies for POC tests. The aim of this study was to establish and validate a LAMP assay applying lyophilized reagents which are stable at ambient temperature, thus avoiding the need for cold-chains. The results from this study suggest that the assay provides a valuable alternative to other PCR tests as currently used for laboratory confirmation of BUD.

Conclusions / Significance: 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.