{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,5]],"date-time":"2026-03-05T20:01:06Z","timestamp":1772740866100,"version":"3.50.1"},"reference-count":56,"publisher":"American Society of Tropical Medicine and Hygiene","issue":"3","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,3,4]]},"abstract":"<jats:title>ABSTRACT.<\/jats:title>\n                  <jats:p>A room temperature-stable and ready-to-use dried-format loop-mediated isothermal amplification (LAMP) diagnostic tool is proposed for detecting knowlesi malaria in the present study. Traditional LAMP assays require a cold chain to maintain the stability of their reagents during storage and transport. This requirement limits the use of LAMP-based methods in environments with limited infrastructure. Results can be obtained by directly observing color changes without the need for specialized instruments; positive samples appear yellow, whereas negative reactions remain pink. The developed dehydrated colorimetric LAMP was tested using 75 clinical blood samples collected from district and state hospitals in Malaysia. By using nested polymerase chain reaction-based assays as a reference method, dehydrated LAMP exhibited 100% sensitivity (95% CI: 92.1\u2013100.0%) and 100% specificity (95% CI: 88.4\u2013100.0%). The dehydrated LAMP reagents can be stored at room temperature for up to 15 days. This dehydrated LAMP assay offers an ideal diagnostic tool for resource-limited settings where knowlesi malaria is endemic.<\/jats:p>","DOI":"10.4269\/ajtmh.25-0201","type":"journal-article","created":{"date-parts":[[2025,12,16]],"date-time":"2025-12-16T14:28:08Z","timestamp":1765895288000},"page":"453-457","source":"Crossref","is-referenced-by-count":0,"title":["Rapid Detection of Knowlesi Malaria Using Room-Temperature Stable and Ready-to-Use Colorimetric Loop-Mediated Isothermal Amplification Reagents"],"prefix":"10.4269","volume":"114","author":[{"given":"Meng Yee","family":"Lai","sequence":"first","affiliation":[{"name":"Department of Parasitology, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yee Ling","family":"Lau","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2795","reference":[{"key":"b1","article-title":"World Malaria Report 2023.","year":"2023"},{"key":"b2","series-title":"Parasit Vectors","first-page":"26","article-title":"Plasmodium knowlesi in humans, macaques and mosquitoes in peninsular Malaysia","volume":"1","author":"Vythilingam","year":"2008"},{"key":"b3","series-title":"Clin Microbiol Rev","first-page":"165","article-title":"Human infections and detection of Plasmodium knowlesi.","volume":"26","author":"Singh","year":"2013"},{"key":"b4","series-title":"PLoS Negl Trop Dis","first-page":"e2026","article-title":"Increasing incidence of Plasmodium knowlesi malaria following control of P. falciparum and P. vivax malaria in Sabah, Malaysia","volume":"7","author":"William","year":"2013"},{"key":"b5","series-title":"Lancet Infect Dis","first-page":"582","article-title":"The reliability of diagnostic techniques in the diagnosis and management of malaria in the absence of a gold standard","volume":"6","author":"Ochola","year":"2006"},{"key":"b6","series-title":"Malar J","first-page":"284","article-title":"Performance of microscopy and RDTs in the context of a malaria prevalence survey in Angola: A comparison using PCR as the gold standard","volume":"12","author":"Fan\u00e7ony","year":"2013"},{"key":"b7","series-title":"Malar J","first-page":"341","article-title":"Significance of nested PCR testing for the detection of low-density malaria infection amongst febrile patients from the Malaria Elimination Demonstration Project in Mandla, Madhya Pradesh, India","volume":"21","author":"Singh","year":"2022"},{"key":"b8","series-title":"Trop Med Infect Dis","first-page":"102","article-title":"Evaluation of malaria diagnostic methods as a key for successful control and elimination programs","volume":"5","author":"Mbanefo","year":"2020"},{"key":"b9","series-title":"Am J Trop Med Hyg","first-page":"119","article-title":"A review of malaria diagnostic tools: Microscopy and rapid diagnostic test (RDT)","volume":"77","author":"Wongsrichanalai","year":"2007"},{"key":"b10","series-title":"Malar J","first-page":"412","article-title":"Accuracy of an HRP-2\/panLDH rapid diagnostic test to detect peripheral and placental Plasmodium falciparum infection in Papua New Guinean women with anaemia or suspected malaria","volume":"14","author":"Umbers","year":"2015"},{"key":"b11","series-title":"Malar J","first-page":"211","article-title":"Loop-mediated isothermal amplification (LAMP) test for diagnosis of uncomplicated malaria in endemic areas: A meta-analysis of diagnostic test accuracy","volume":"19","author":"Selvarajah","year":"2020"},{"key":"b12","series-title":"Front Cell Infect Microbiol","first-page":"961832","article-title":"Rapid diagnosis of Plasmodium falciparum malaria using a point of-care loop-mediated isothermal amplification device","volume":"12","author":"Puri","year":"2022"},{"key":"b13","series-title":"J Infect Dis","first-page":"1167","article-title":"Point of care testing for malaria using LAMP, loop mediated isothermal amplification","volume":"210","author":"Hsiang","year":"2014"},{"key":"b14","series-title":"Malar J","first-page":"166","article-title":"Validation of SYBR green I based closed-tube loop-mediated isothermal amplification (LAMP) assay for diagnosis of knowlesi malaria","volume":"20","author":"Lai","year":"2021"},{"key":"b15","series-title":"J Clin Microbiol","first-page":"4173","article-title":"Spurious amplification of a Plasmodium vivax small-subunit RNA gene by use of primers currently used to detect P. knowlesi.","volume":"47","author":"Imwong","year":"2009"},{"key":"b16","series-title":"Mol Biochem Parasitol","first-page":"315","article-title":"High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction","volume":"61","author":"Snounou","year":"1993"},{"key":"b17","series-title":"Am J Trop Med Hyg","first-page":"336","article-title":"Loop-mediated isothermal amplification assay for identification of five human Plasmodium species in Malaysia","volume":"94","author":"Lau","year":"2016"},{"key":"b18","article-title":"Diagnostic Test Evaluation Calculator.","year":"2025"},{"key":"b19","series-title":"Am J Trop Med Hyg","first-page":"1370","article-title":"Evaluation of WarmStart colorimetric loop-mediated isothermal amplification assay for diagnosis of malaria","volume":"102","author":"Lai","year":"2020"},{"key":"b20","series-title":"Anal Chim Acta","first-page":"71","article-title":"Development of a thermostabilised triplex LAMP assay with dry-reagent four target lateral flow dipstick for detection of Entamoeba histolytica and non-pathogenic Entamoeba spp","volume":"966","author":"Foo","year":"2017"},{"key":"b21","series-title":"J Microbiol Methods","first-page":"99","article-title":"Development of a dry-reagent-based nucleic acid-sensing platform by coupling thermostabilised LATE-PCR assay to an oligonucleotide-modified lateral flow biosensor","volume":"118","author":"Ang","year":"2015"},{"key":"b22","series-title":"Biophys Chem","first-page":"25","article-title":"The thermodynamic mechanism of protein stabilization by trehalose","volume":"64","author":"Xie","year":"1997"},{"key":"b23","series-title":"BMC Microbiol","first-page":"36","article-title":"A novel ready-to-use loop-mediated isothermal amplification (LAMP) method for detection of Burkholderia mallei and B. pseudomallei.","volume":"25","author":"Nakase","year":"2025"},{"key":"b24","series-title":"PLoS Negl Trop Dis","first-page":"e0003578","article-title":"Direct blood dry LAMP: A rapid, stable, and easy diagnostic tool for human African Trypanosomiasis","volume":"9","author":"Hayashida","year":"2015"},{"key":"b25","series-title":"Tuberculosis (Edinb)","first-page":"1","article-title":"Direct detection of Mycobacterium tuberculosis in clinical samples by a dry methyl green loop-mediated isothermal amplification (LAMP) method","volume":"117","author":"Thapa","year":"2019"},{"key":"b26","series-title":"J Vet Diagn Invest","first-page":"307","article-title":"Direct detection of Mycobacterium bovis by a dry loop-mediated isothermal amplification assay in cattle samples collected during routine abattoir examination in Malawi","volume":"35","author":"Kapalamula","year":"2023"},{"key":"b27","series-title":"Sci Rep","first-page":"7043","article-title":"A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogens","volume":"12","author":"Song","year":"2022"},{"key":"b28","series-title":"Asian Pac J Trop Med","first-page":"428","article-title":"A simple and purification-free nucleic acid extraction method for rapid diagnosis of malaria","volume":"16","author":"Lee","year":"2023"},{"key":"b1","article-title":"World Malaria Report 2023.","year":"2023"},{"key":"b2","series-title":"Parasit 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malaria prevalence survey in Angola: A comparison using PCR as the gold standard","volume":"12","author":"Fan\u00e7ony","year":"2013"},{"key":"b7","series-title":"Malar J","first-page":"341","article-title":"Significance of nested PCR testing for the detection of low-density malaria infection amongst febrile patients from the Malaria Elimination Demonstration Project in Mandla, Madhya Pradesh, India","volume":"21","author":"Singh","year":"2022"},{"key":"b8","series-title":"Trop Med Infect Dis","first-page":"102","article-title":"Evaluation of malaria diagnostic methods as a key for successful control and elimination programs","volume":"5","author":"Mbanefo","year":"2020"},{"key":"b9","series-title":"Am J Trop Med Hyg","first-page":"119","article-title":"A review of malaria diagnostic tools: Microscopy and rapid diagnostic test (RDT)","volume":"77","author":"Wongsrichanalai","year":"2007"},{"key":"b10","series-title":"Malar J","first-page":"412","article-title":"Accuracy of an HRP-2\/panLDH rapid diagnostic test to detect peripheral and placental Plasmodium falciparum infection in Papua New Guinean women with anaemia or suspected malaria","volume":"14","author":"Umbers","year":"2015"},{"key":"b11","series-title":"Malar J","first-page":"211","article-title":"Loop-mediated isothermal amplification (LAMP) test for diagnosis of uncomplicated malaria in endemic areas: A meta-analysis of diagnostic test accuracy","volume":"19","author":"Selvarajah","year":"2020"},{"key":"b12","series-title":"Front Cell Infect Microbiol","first-page":"961832","article-title":"Rapid diagnosis of Plasmodium falciparum malaria using a point of-care loop-mediated isothermal amplification device","volume":"12","author":"Puri","year":"2022"},{"key":"b13","series-title":"J Infect Dis","first-page":"1167","article-title":"Point of care testing for malaria using LAMP, loop mediated isothermal amplification","volume":"210","author":"Hsiang","year":"2014"},{"key":"b14","series-title":"Malar J","first-page":"166","article-title":"Validation of SYBR green I based closed-tube loop-mediated isothermal amplification (LAMP) assay for diagnosis of knowlesi malaria","volume":"20","author":"Lai","year":"2021"},{"key":"b15","series-title":"J Clin Microbiol","first-page":"4173","article-title":"Spurious amplification of a Plasmodium vivax small-subunit RNA gene by use of primers currently used to detect P. knowlesi.","volume":"47","author":"Imwong","year":"2009"},{"key":"b16","series-title":"Mol Biochem Parasitol","first-page":"315","article-title":"High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction","volume":"61","author":"Snounou","year":"1993"},{"key":"b17","series-title":"Am J Trop Med Hyg","first-page":"336","article-title":"Loop-mediated isothermal amplification assay for identification of five human Plasmodium species in Malaysia","volume":"94","author":"Lau","year":"2016"},{"key":"b18","article-title":"Diagnostic Test 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stabilization by trehalose","volume":"64","author":"Xie","year":"1997"},{"key":"b23","series-title":"BMC Microbiol","first-page":"36","article-title":"A novel ready-to-use loop-mediated isothermal amplification (LAMP) method for detection of Burkholderia mallei and B. pseudomallei.","volume":"25","author":"Nakase","year":"2025"},{"key":"b24","series-title":"PLoS Negl Trop Dis","first-page":"e0003578","article-title":"Direct blood dry LAMP: A rapid, stable, and easy diagnostic tool for human African Trypanosomiasis","volume":"9","author":"Hayashida","year":"2015"},{"key":"b25","series-title":"Tuberculosis (Edinb)","first-page":"1","article-title":"Direct detection of Mycobacterium tuberculosis in clinical samples by a dry methyl green loop-mediated isothermal amplification (LAMP) method","volume":"117","author":"Thapa","year":"2019"},{"key":"b26","series-title":"J Vet Diagn Invest","first-page":"307","article-title":"Direct detection of Mycobacterium bovis by a dry loop-mediated 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