LAMP Assay for the Diagnosis of Visceral Leishmaniasis (EvaLAMP)

Evaluation of the Loop-mediated Amplification Assay and Direct-Blood PCR-Nucleic-Acid Lateral Flow Immuno-Assay for the Diagnosis and/or as Test-of-Cure in Patients With Visceral Leishmaniasis in Ethiopia

This study will evaluate the of the loop-mediated amplification assay (LAMP) as a diagnostic as well as a Test-of-Cure (ToC) for visceral leishmaniasis (VL) in an endemic area in Ethiopia. Furthermore, we aim to further development of the direct-blood PCR-Nucleic Acid Lateral-Flow Immuno-Assay (dB-PCR-NALFIA) as a novel diagnostic tool for VL and its subsequent evaluation in the field.

Study Overview

• Status: Completed
• Conditions: Visceral Leishmaniasis

Detailed Description

Leishmaniasis is among the World's important neglected infectious diseases (NIDs). The WHO estimates that 350 million people are at risk of contracting leishmaniasis. Visceral leishmaniasis (VL) is the most severe form of the disease. Ethiopia has been recently listed by WHO among the fourteen countries in the world with the highest burden of VL. The development of novel point-of-care (PoC) diagnostics and/or a Test-of-Cure (ToC) for VL is deemed a key priority research area. In several endemic areas current gold standard diagnosis and monitoring of treatment efficacy of VL is based on parasite detection or serology. However, these tests are either not available for routine use or lack sufficient sensitivity and specificity, in particular in HIV co-infected patients. Though molecular tests such as PCR have become popular choices as a tool to diagnose VL, monitor treatment response and predict relapse, these techniques require technical skill and equipment and are considerably more expensive. Recent advances in diagnostics has been the development of LAMP with several advantages, such as no need for thermocycler, high specificity, simple read-out and no cold chain requirements. Therefore, LAMP has emerged as a powerful tool for PoC diagnostics. Its clinical utility as PoC diagnosis and/or ToC for VL in the African setting is, however, hardly known.

Here, the investigators will evaluate the utility of the LAMP as a PoC and/or ToC for VL in an endemic area in Ethiopia. The performance of the LAMP assay as a diagnostic tool will be evaluated in newly diagnosed VL cases confirmed by parasite detection and/or PCR. Furthermore, the use of the assay as ToC will be determined by evaluating the performance of the assay in VL patients confirmed cured at day 17 of therapy, as assessed by negative parasite and/or PCR results. Additionally, the investigators plan to utilize a newly developed rapid molecular platform, db-PCR-NALFIA, which does not require DNA extraction, has an internal amplification control and simple read-out. The investigators will evaluate the utility of both assays also in patients co-infected with HIV. The results may have major policy implications as the application represents a concept that could enhance evidence- based translation of research to improve public health practice by contributing to leishmaniasis management guidelines - with overarching impacts for National, Regional and Global programs.

• Study Type: Observational
• Enrollment (Actual): 500

Contacts and Locations

• Study Contact: Name: Dawit Wolday, MD, PhD; Phone Number: +251911208984; Email: dawwol@gmail.com
• Study Contact Backup: Name: Yazezew Kebede, MD; Phone Number: +251910104423; Email: yazezew@gmail.com

Study Locations

• Ethiopia
  • Mekele, Ethiopia
    • Mekelle University College of Health Sciences

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 3 years to 88 years (Child, Adult, Older Adult)
• Accepts Healthy Volunteers: N/A

• Sampling Method: Non-Probability Sample

Study Population

VL suspected cases will be recruited from Ayder Referral Hospital, Mekelle University College of Health Sciences - (MUCHS) in Mekelle City, the capital of Tigray Regional State in Northern Ethiopia, where VL is highly endemic in the area. Patients seeking care at the hospital will be eligible and screened for inclusion in the study.

Description

Inclusion Criteria:

• Clinical evidence consistent with VL confirmed by microscopy (+ culture) and/or PCR

Exclusion Criteria:

• Treatment with any anti-leishmanial drugs within the previous 3 months
• Not capable of understanding or complying with the study protocol
• Refusal to consent and participate in to the study

Study Plan

How is the study designed?

Design Details

• Observational Models: Cohort
• Time Perspectives: Prospective

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Number of participants correctly diagnosed with VL as assessed by LAMP assay	Performance of the LAMP will be compared to gold- standard diagnostic procedures, including parasite detection and/or PCR-technology	Baseline
Number of participants treated for VL and identified as cured (ToC) at day 17 post-treatment based on the assessment by LAMP assay	LAMP will be compared to gold- standard diagnostic procedures, including parasite detection and/or PCR-technology	Baseline

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Number of participants co-infected with HIV correctly diagnosed with VL as well as treated participants identified as cured (ToC) at day 17 based on the assessment by LAMP assay	LAMP will be compared to gold-standard diagnostic procedures, including parasite detection and/or PCR-technology	17 days
Number of participants correctly diagnosed as VL based on db-PCR-NALFIA technology	The investigators will develop db-PCR-NALFIA technology for the diagnosis of VL, i.e. sample preparation and result read-out will be adopted using db-PCR-NALFIA technology as PoC platform. It's performance will be evaluated against gold-standard.	Baseline

Collaborators and Investigators

• Sponsor: Prof. Dawit Wolday
• Collaborators: Academisch Medisch Centrum - Universiteit van Amsterdam (AMC-UvA); European and Developing Countries Clinical Trials Partnership (EDCTP)
• Investigators: Study Chair: Amanuel Haile, MD, Mekelle University College of Health Sciences

Publications and helpful links

General Publications

• Hagos DG, Schallig HDFH, Kiros YK, Abdulkadir M, Wolday D. Performance of rapid rk39 tests for the diagnosis of visceral leishmaniasis in Ethiopia: a systematic review and meta-analysis. BMC Infect Dis. 2021 Nov 17;21(1):1166. doi: 10.1186/s12879-021-06826-w.
• Hagos DG, Kiros YK, Abdulkader M, Arefaine ZG, Nigus E, Schallig HHDF, Wolday D. Utility of the Loop-Mediated Isothermal Amplification Assay for the Diagnosis of Visceral Leishmaniasis from Blood Samples in Ethiopia. Am J Trop Med Hyg. 2021 Jul 26;105(4):1050-1055. doi: 10.4269/ajtmh.21-0334.
• Hagos DG, Kebede Y, Abdulkader M, Nigus E, Gessesse Arefaine Z, Nega G, Schallig HDF, Wolday D. Effect of rK39 testing in guiding treatment initiation and outcome in patients with visceral leishmaniasis in Ethiopia: A prospective cohort study. PLoS One. 2021 Jun 14;16(6):e0253303. doi: 10.1371/journal.pone.0253303. eCollection 2021.

Study record dates

Study Major Dates

• Study Start (Actual): October 1, 2018
• Primary Completion (Actual): June 30, 2022
• Study Completion (Actual): June 30, 2023

Study Registration Dates

• First Submitted: June 27, 2019
• First Submitted That Met QC Criteria: June 28, 2019
• First Posted (Actual): July 1, 2019

Study Record Updates

• Last Update Posted (Actual): December 20, 2023
• Last Update Submitted That Met QC Criteria: December 14, 2023
• Last Verified: December 1, 2023

More Information

Terms related to this study

• Keywords: PCR; Diagnosis; HIV co-infection; Test-of-Cure; Loop-mediated amplification assay; Nucleic-acid Lateral Flow Immunoassay
• Additional Relevant MeSH Terms: Skin Diseases; Infections; Vector Borne Diseases; Parasitic Diseases; Protozoan Infections; Skin Diseases, Parasitic; Skin Diseases, Infectious; Euglenozoa Infections; Leishmaniasis; Leishmaniasis, Visceral
• Other Study ID Numbers: TMA2016SF-1437

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES
• IPD Plan Description: De-identified individual patient data for all primary and secondary outcome measures will be made available to anyone who submit requests.
• IPD Sharing Time Frame: After 12 months of completion of project
• IPD Sharing Access Criteria: Data access requests will be first reviewed by the project's steering committee to ensure that all requested analyses can be achieved with the available study data.
• IPD Sharing Supporting Information Type: STUDY_PROTOCOL; SAP; ICF; ANALYTIC_CODE; CSR

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No