Abstract
Elimination of Human African Trypanosomiasis (HAT) requires highly specific and sensitive tools for both diagnostic at point of care and epidemiological surveys. We have adapted SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) for the detection of trypanosome nucleic acids. Our SHERLOCK4HAT diagnostic tool kit, using 7SLRNA, TgSGP and SRA targets, distinguishes between Trypanosoma brucei (T. b.) brucei, T. b. gambiense (g) and T. b. rhodesiense (r) without cross-reactivity and with sensitivity between 0.01 and 0.1 parasite/µL. SHERLOCK4HAT can accurately detect a trypanosome infection in cryo-banked patient buffy coats, with 85.1% sensitivity and 98.4% specificity for gHAT, and 100% sensitivity and 94.1% specificity for rHAT. Our SHERLOCK4HAT diagnostic showed 85.6% correlation with a reference standard qPCR in gHAT patients, 96.2% correlation in rHAT patients, discriminates between r/gHAT with 100% accuracy and is compatible with lateral flow assay readout for use at the point of care.
Competing Interest Statement
The authors have declared no competing interest.
Funding Statement
This project has received a funding from the Institut Pasteur to BR and LG (PTR-175 SHERLOCK4HAT) and from the French Governments Investissement d Avenir program Laboratoire d Excellence Integrative Biology of Emerging Infectious Diseases (LabEx IBEID). NS was supported by funding from the Institut Pasteur (PTR-175 SHERLOCK4HAT). The funders had no role in study design or data collection and analysis or decision to publish or preparation of the manuscript.
Author Declarations
I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained.
Yes
The details of the IRB/oversight body that provided approval or exemption for the research described are given below:
Blood from healthy human donors was provided by ICAReB platform (Clinical Investigation & Access to Research Bioresources) in the Center for Translational Science, at the Institut Pasteur (Paris). All participants gave written informed consent in the frame of the healthy volunteers Diagmicoll cohort (Clinical trials NCT 03912246) after approval of the CPP Ile-de-France I Ethics Committee (2009, April 30th). For the WHO HAT Specimen biobank samples, approval was given by the WHO Ethical Review Committee, each national ethical committee where samples were taken and the national Ministries of Health.
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Footnotes
↵† Co-last authors
Data Availability
All data are available in the main text or the supplementary materials.