Diagnostic performance of a Recombinant Polymerase Amplification Test-Lateral Flow (RPA-LF) for cutaneous leishmaniasis in an endemic setting of Colombia

Alexandra Cossio, Jimena Jojoa, María Del Mar Castro, Ruth Mabel Castillo, Lyda Osorio, Thomas R Shelite, Nancy Gore Saravia, Peter C Melby, Bruno L Travi, Alexandra Cossio, Jimena Jojoa, María Del Mar Castro, Ruth Mabel Castillo, Lyda Osorio, Thomas R Shelite, Nancy Gore Saravia, Peter C Melby, Bruno L Travi

Abstract

Background: Control of cutaneous leishmaniasis by public health systems in the Americas relies on case identification and treatment. Point-of-care diagnostics that can be performed by health workers within or near affected communities could effectively bring the health system to the resource-limited sites providing early diagnosis and treatment, reducing morbidity and the burden of disease.

Methodology/principal findings: A cross-sectional study was undertaken to evaluate the diagnostic test performance of Isothermal Recombinase Polymerase Amplification (RPA) targeting Leishmania kinetoplast DNA, coupled with a lateral flow (LF) immunochromatographic strip, in a field setting and a laboratory reference center. Minimally invasive swab and FTA filter paper samples were obtained by community health workers and highly trained technicians from ulcerated lesions of > 2 weeks' evolution from 118 patients' ≥ 2 years of age in the municipality of Tumaco, Nariño. Extracted DNA was processed by RPA-LF at a reference center or in a primary health facility in the field. Evaluation was based on a composite "gold standard" that included microscopy, culture, biopsy and real-time polymerase chain reaction detection of Leishmania 18S rDNA. Standard of care routine diagnostic tests were explored as comparators. Sensitivity and specificity of RPA-LF in the reference lab scenario were 87% (95%CI 74-94) and 86% (95%CI 74-97), respectively. In the field scenario, the sensitivity was 75% (95%CI 65-84) and specificity 89% (95%CI 78-99). Positive likelihood ratios in both scenarios were higher than 6 while negative likelihood ratios ranged to 0.2-0.3 supporting the usefulness of RPA-LF to rule-in and potentially to rule-out infection.

Conclusions/significance: The low complexity requirements of RPA-LF combined with non-invasive sampling support the feasibility of its utilization by community health workers with the goal of strengthening the diagnostic capacity for cutaneous leishmaniasis in Colombia.

Trial registration: ClinicalTrials.gov NCT04500873.

Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1. Schematic summary of patient enrollment,…
Fig 1. Schematic summary of patient enrollment, and subsequent, sampling, diagnostic procedures, and performance of RPA-LF evaluation in two scenarios.
A) Reference lab scenario: samples were obtained by highly trained field technician in Tumaco and processed in a reference center in Cali. In Cali samples were obtained and processed by an expert microbiologist. B) Field scenario: samples were obtained by trained community health workers (CHW), RPA-LF was processed by a non-expert field technician in Tumaco in primary health facility (PHF), and swabs were sent to reference center to be processed. C. Performance of RPA-LF test.
Fig 2. Results of RPA-LF test
Fig 2. Results of RPA-LF test
Fig 3. Flow chart of participants by…
Fig 3. Flow chart of participants by scenario.

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