{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T16:08:45Z","timestamp":1791216525683,"version":"4.1.0"},"reference-count":32,"publisher":"American Society of Tropical Medicine and Hygiene","issue":"4","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,4,3]]},"abstract":"<jats:title>ABSTRACT.<\/jats:title>\n<jats:p>Loop-mediated isothermal amplification (LAMP) is a nucleic acid amplification technique that can amplify specific nucleic acids at a constant temperature (63\u201365\u00b0C) within a short period (&lt;1\u2009hour). In this study, we report the utilization of recombinase-aided LAMP to specifically amplify the <jats:italic>18S sRNA<\/jats:italic> of <jats:italic>Plasmodium knowlesi.<\/jats:italic> The method was built on a conventional LAMP assay by inclusion of an extra enzyme, namely recombinase, into the master mixture. With the addition of recombinase into the LAMP assay, the assay speed was executed within a time frame of less than 28\u2009minutes at 65\u00b0C. We screened <jats:italic>55\u2009P. knowlesi<\/jats:italic> samples and 47 non\u2013<jats:italic>P. knowlesi<\/jats:italic> samples. No cross-reactivity was observed for non\u2013<jats:italic>P. knowlesi<\/jats:italic> samples, and the detection limit for recombinase-aided LAMP was one copy for <jats:italic>P. knowlesi<\/jats:italic> after LAMP amplification. It has been reported elsewhere that LAMP can be detected through fluorescent readout systems. Although such systems result in considerable limits of detection, the need for sophisticated equipment limits their use. Hence, we used here a colorimetric detection platform for the evaluation of the LAMP assay\u2019s performance. This malachite green\u2013based recombinase-aided LAMP assay enabled visualization of results with the naked eye. Negative samples were observed by a change in color from green to colorless, whereas positive samples remained green. Our results demonstrate that the LAMP assay developed here is a convenient, sensitive, and useful diagnostic tool for the rapid detection of knowlesi malaria parasites. This method is suitable for implementation in remote healthcare settings, where centralized laboratory facilities, funds, and clinicians are in short supply.<\/jats:p>","DOI":"10.4269\/ajtmh.23-0572","type":"journal-article","created":{"date-parts":[[2024,2,27]],"date-time":"2024-02-27T18:35:09Z","timestamp":1709058909000},"page":"648-652","source":"Crossref","is-referenced-by-count":3,"title":["Recombinase-Aided Loop-Mediated Isothermal Amplification on Human Plasmodium knowlesi"],"prefix":"10.4269","volume":"110","author":[{"given":"Meng Yee","family":"Lai","sequence":"first","affiliation":[{"name":"Department of Parasitology, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, Malaysia;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohd Hafizi","family":"Abdul Hamid","sequence":"additional","affiliation":[{"name":"Vector Borne Disease Sector, Ministry of Health, Putrajaya, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jenarun","family":"Jelip","sequence":"additional","affiliation":[{"name":"Vector Borne Disease Sector, Ministry of Health, Putrajaya, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rose Nani","family":"Mudin","sequence":"additional","affiliation":[{"name":"Vector Borne Disease Sector, Ministry of Health, Putrajaya, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yee Ling","family":"Lau","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, Malaysia;"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2795","reference":[{"key":"b1","series-title":"Clin Microbiol Infect","first-page":"640","article-title":"Human infections with Plasmodium knowlesi-zoonotic malaria","volume":"21","author":"Millar","year":"2015"},{"key":"b2","series-title":"World Malaria Report 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One","first-page":"e0258722","article-title":"Validation study of boil & spin malachite green loop mediated isothermal amplification (B&S MG LAMP) versus microscopy for malaria detection in the Peruvian Amazon","volume":"16","author":"Barazorda","year":"2021"},{"key":"b27","series-title":"J Mol Diagn","first-page":"729","article-title":"Development of reverse transcription loop-mediated isothermal amplification assays targeting severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2)","volume":"22","author":"Park","year":"2020"},{"key":"b28","series-title":"PLoS One","first-page":"e0151437","article-title":"Use of malachite green-loop-mediated isothermal amplification for detection of Plasmodium spp. parasites","volume":"11","author":"Lucchi","year":"2016"},{"key":"b29","series-title":"PLoS One","first-page":"e0235216","article-title":"Evaluation and improvement of isothermal amplification methods for point-of-need plant disease 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