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Performance evaluation of the diaxxoPCR system for rapid and user-friendly stool-based diagnosis of trichuriasis, ascariasis and strongyloidiasis in Mozambique
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Resumo(s)
BACKGROUND: Nucleic acid amplification tests have shown promising results for the diagnosis of soil-transmitted helminths (STH). The implementation of real-time PCR (qPCR) in low-resource settings is, however, still hampered by multiple logistical challenges. In this study, we assessed the diagnostic performance of a cartridge-based real-time PCR system (diaxxoPCR) for the detection of DNA of Trichuris trichiura, Ascaris lumbricoides, and Strongyloides stercoralis in clinical samples, using qPCR as the reference test. METHODOLOGY: Initially, a technical validation of the diaxxoPCR system in a singleplex pod (cartridge) design was performed using 37 predefined DNA samples (Study A), followed by a diagnostic comparison between the diaxxoPCR system (singleplex) and qPCR on DNA samples extracted from 325 stools collected in a clinical trial in a rural area of Mozambique (Study B). Finally, one negative and one positive DNA sample were used to demonstrate the technical performance of a multiplex pod design as a potential use-case for the diaxxoPCR system (Study C). PRINCIPAL FINDINGS: Study A demonstrated that the diaxxoPCR system performed reliably for each of the three STH targets, with minimal intra- and inter-assay variation and sufficient output reproducibility. Study B, performed in Mozambique, showed a positive qPCR result in 57.5% (187), 15.4% (50), and 0.3% (1) of the 325 DNA trial samples for T. trichiura, A. lumbricoides, and S. stercoralis, respectively. The diaxxoPCR system demonstrated sensitivities and specificities above 97% and 94% for each target, resulting in nearly perfect to perfect qualitative agreements with the reference test. Quantitatively, significant and positive associations were seen between the Ct-values (qPCR) and Cq-values (diaxxoPCR). In Study C, the diaxxoPCR system correctly detected all 3 targets in the multiplex pod design. CONCLUSION: With refinements regarding faecal DNA extraction procedures, the diaxxoPCR system has potential to provide accurate and easy-to-use real-time molecular diagnostics of STH in low resource laboratories.
Descrição
Funding text: This project was funded at CISM and the LUMC by the EDCTP2 program supported by the European Union (grant number RIA2017NCT-1845 -STOP; www.stoptheworm.org) Horizon 2020 European Union Funding for Research and Innovation (JM, LvL, AK and IM). The funders had no role in the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: Copyright: © 2025 Messa et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Palavras-chave
Public Health, Environmental and Occupational Health Infectious Diseases SDG 3 - Good Health and Well-being