Diagnostic accuracy of loopamp Trypanosoma brucei detection kit for diagnosis of human African trypanosomiasis in clinical samples

Patrick Mitashi, Epco Hasker, Dieudonné Mumba Ngoyi, Pati Patient Pyana, Veerle Lejon, Wim Van der Veken, Pascal Lutumba, Philippe Büscher, Marleen Boelaert, Stijn Deborggraeve, Patrick Mitashi, Epco Hasker, Dieudonné Mumba Ngoyi, Pati Patient Pyana, Veerle Lejon, Wim Van der Veken, Pascal Lutumba, Philippe Büscher, Marleen Boelaert, Stijn Deborggraeve

Abstract

Background: Molecular methods have great potential for sensitive parasite detection in the diagnosis of human African trypanosomiasis (HAT), but the requirements in terms of laboratory infrastructure limit their use to reference centres. A recently developed assay detects the Trypanozoon repetitive insertion mobile element (RIME) DNA under isothermal amplification conditions and has been transformed into a ready-to-use kit format, the Loopamp Trypanosoma brucei. In this study, we have evaluated the diagnostic performance of the Loopamp Trypanosoma brucei assay (hereafter called LAMP) in confirmed T.b. gambiense HAT patients, HAT suspects and healthy endemic controls from the Democratic Republic of the Congo (DRC).

Methodology/principal findings: 142 T.b. gambiense HAT patients, 111 healthy endemic controls and 97 HAT suspects with unconfirmed status were included in this retrospective evaluation. Reference standard tests were parasite detection in blood, lymph or cerebrospinal fluid. Archived DNA from blood of all study participants was analysed in duplicate with LAMP. Sensitivity of LAMP in parasitologically confirmed cases was 87.3% (95% CI 80.9-91.8%) in the first run and 93.0% (95% CI 87.5-96.1%) in the second run. Specificity in healthy controls was 92.8% (95% CI 86.4-96.3%) in the first run and 96.4% (95% CI 91.1-98.6%) in the second run. Reproducibility was excellent with a kappa value of 0.81.

Conclusions/significance: In this laboratory-based study, the Loopamp Trypanosoma brucei Detection Kit showed good diagnostic accuracy and excellent reproducibility. Further studies are needed to assess the feasibility of its routine use for diagnosis of HAT under field conditions.

Conflict of interest statement

I have read the journal's policy and have the following conflicts: FIND, the sponsor of the development of the LAMP kit, made useful suggestions on the technical protocol and provided the required kits and reagents for this study. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. This does not alter our adherence to all PLOS policies on sharing data and materials.

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