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However, both techniques have their constraints, including poor sensitivity for low parasitaemias. Hence, more accurate diagnostic tests for field use and routine clinical settings are warranted. The miniature direct-on-blood PCR nucleic acid lateral flow immunoassay (mini-dbPCR-NALFIA) is an innovative, easy-to-use molecular assay for diagnosis of malaria in resource-limited settings. Unlike traditional molecular methods, mini-dbPCR-NALFIA does not require DNA extraction and makes use of a handheld, portable thermal cycler that can run on a solar-charged power pack. Result read-out is done using a rapid lateral flow strip enabling differentiation of <jats:italic>Plasmodium falciparum<\/jats:italic> and non-<jats:italic>falciparum<\/jats:italic> malaria infections. A laboratory evaluation was performed to assess the performance of the mini-dbPCR-NALFIA for diagnosis of pan-<jats:italic>Plasmodium<\/jats:italic> and <jats:italic>P. falciparum<\/jats:italic> infections in whole blood.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>Diagnostic accuracy of the mini-dbPCR-NALFIA was determined by testing a set of <jats:italic>Plasmodium<\/jats:italic>-positive blood samples from returned travellers (n\u2009=\u200929), and <jats:italic>Plasmodium<\/jats:italic>-negative blood samples from travellers with suspected malaria (n\u2009=\u200923), the Dutch Blood Bank (n\u2009=\u200919) and intensive care patients at the Amsterdam University Medical Centers (n\u2009=\u200916). Alethia Malaria (LAMP) with microscopy for species differentiation were used as reference. Limit of detection for <jats:italic>P. falciparum<\/jats:italic> was determined by 23 measurements of a dilution series of a <jats:italic>P. falciparum<\/jats:italic> culture. A fixed sample set was tested three times by the same operator to evaluate the repeatability, and once by five different operators to assess the reproducibility.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Overall sensitivity and specificity of the mini-dbPCR-NALFIA were 96.6% (95% CI, 82.2%\u201399.9%) and 98.3% (95% CI, 90.8%\u2013100%). Limit of detection for <jats:italic>P. falciparum<\/jats:italic> was 10 parasites per microlitre of blood. The repeatability of the assay was 93.7% (95% CI, 89.5%\u201397.8%) and reproducibility was 84.6% (95% CI, 79.5%\u201389.6%).<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>Mini-dbPCR-NALFIA is a sensitive, specific and robust method for molecular diagnosis of <jats:italic>Plasmodium<\/jats:italic> infections in whole blood and differentiation of <jats:italic>P. falciparum<\/jats:italic>. Incorporation of a miniature thermal cycler makes the assay well-adapted to resource-limited settings. A phase-3 field trial is currently being conducted to evaluate the potential implementation of this tool in different malaria transmission areas.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s12936-023-04496-4","type":"journal-article","created":{"date-parts":[[2023,3,17]],"date-time":"2023-03-17T13:02:59Z","timestamp":1679058179000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Laboratory evaluation of the miniature direct-on-blood PCR nucleic acid lateral flow immunoassay (mini-dbPCR-NALFIA), a simplified molecular diagnostic test for Plasmodium"],"prefix":"10.1186","volume":"22","author":[{"given":"Norbert J.","family":"van Dijk","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandra","family":"Menting","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ellen M. 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Informed consent was obtained from human participants for the use of anonymized specimens according to the \u2018no objection\u2019 system, hence the study was exempted from the need for ethical approval according to Dutch law.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"EW is an employee of Abingdon Health, manufacturer of the NALFIA strips used in the present study. Abingdon Health was not directly involved in the design of the study or the data analysis.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"98"}}