{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T12:03:25Z","timestamp":1791461005609,"version":"4.3.4"},"update-to":[{"DOI":"10.1371\/journal.pntd.0012129","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2025,2,19]],"date-time":"2025-02-19T00:00:00Z","timestamp":1739923200000}}],"reference-count":49,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2025,2,6]],"date-time":"2025-02-06T00:00:00Z","timestamp":1738800000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"DOI":"10.13039\/501100000974","name":"Australian Centre for International Agricultural Research","doi-asserted-by":"publisher","award":["LS-2019-116"],"award-info":[{"award-number":["LS-2019-116"]}],"id":[{"id":"10.13039\/501100000974","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000996","name":"Department of Foreign Affairs and Trade, Australian Government","doi-asserted-by":"publisher","award":["LS-2019-116"],"award-info":[{"award-number":["LS-2019-116"]}],"id":[{"id":"10.13039\/501100000996","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01AI160457-01"],"award-info":[{"award-number":["R01AI160457-01"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009898","name":"Defense Health Agency","doi-asserted-by":"publisher","award":["P0097_22_N2"],"award-info":[{"award-number":["P0097_22_N2"]}],"id":[{"id":"10.13039\/100009898","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["1037304"],"award-info":[{"award-number":["1037304"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["1045156"],"award-info":[{"award-number":["1045156"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["1042072"],"award-info":[{"award-number":["1042072"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["2017436"],"award-info":[{"award-number":["2017436"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["2016792"],"award-info":[{"award-number":["2016792"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100013885","name":"Kementerian Kesihatan Malaysia","doi-asserted-by":"publisher","award":["BP00500\/117\/1002"],"award-info":[{"award-number":["BP00500\/117\/1002"]}],"id":[{"id":"10.13039\/501100013885","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["1042072"],"award-info":[{"award-number":["1042072"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"\n Background<\/jats:title>\n \n Zoonotic\n P<\/jats:italic>\n .\n knowlesi<\/jats:italic>\n and\n P<\/jats:italic>\n .\n cynomolgi<\/jats:italic>\n symptomatic and asymptomatic infections occur across endemic areas of Southeast Asia. Most infections are low-parasitemia, with an unknown proportion below routine microscopy detection thresholds. Molecular surveillance tools optimizing the limit of detection (LOD) would allow more accurate estimates of zoonotic malaria prevalence.\n <\/jats:p>\n <\/jats:sec>\n \n Methodology\/Principal findings<\/jats:title>\n \n An established ultra-sensitive\n Plasmodium<\/jats:italic>\n genus quantitative-PCR (qPCR) assay targeting the 18S rRNA gene underwent LOD evaluation with and without reverse transcription (RT) for\n P<\/jats:italic>\n .\n knowlesi<\/jats:italic>\n ,\n P<\/jats:italic>\n .\n cynomolgi<\/jats:italic>\n and\n P<\/jats:italic>\n .\n vivax<\/jats:italic>\n using total nucleic acid preserved (DNA\/RNA Shield) isolates and archived dried blood spots (DBS). LODs for selected\n P<\/jats:italic>\n .\n knowlesi-<\/jats:italic>\n specific assays, and reference\n P<\/jats:italic>\n .\n vivax-<\/jats:italic>\n and\n P<\/jats:italic>\n .\n cynomolgi<\/jats:italic>\n -specific assays were determined with reverse transcription (RT). Assay specificities were assessed using clinical malaria samples and malaria-negative controls.\n <\/jats:p>\n \n The use of reverse transcription improved\n Plasmodium<\/jats:italic>\n species detection by up to 10,000-fold (\n Plasmodium<\/jats:italic>\n genus), 2759-fold (\n P<\/jats:italic>\n .\n knowlesi<\/jats:italic>\n ) and 1000-fold (\n P<\/jats:italic>\n .\n vivax<\/jats:italic>\n and\n P<\/jats:italic>\n .\n cynomolgi<\/jats:italic>\n ).\n <\/jats:p>\n \n The Kamau et al.\n Plasmodium<\/jats:italic>\n genus RT-qPCR assay was highly sensitive for\n P<\/jats:italic>\n .\n knowlesi<\/jats:italic>\n detection with a median LOD of \u22640.0002 parasites\/\u03bcL compared to 0.002 parasites\/\u03bcL for\n P<\/jats:italic>\n .\n cynomolgi<\/jats:italic>\n and\n P<\/jats:italic>\n .\n vivax<\/jats:italic>\n . The LODs with RT for\n P<\/jats:italic>\n .\n knowlesi<\/jats:italic>\n -specific PCRs were enhanced for the Imwong et al. 18S rRNA (0.0007 parasites\/\u03bcL) and Divis et al. real-time 18S rRNA (0.0002 parasites\/\u03bcL) assays, but not for the Lubis et al. hemi-nested\n SICAvar<\/jats:italic>\n (1.1 parasites\/\u03bcL) and Lee et al. nested 18S rRNA (11 parasites\/\u03bcL). The LOD for\n P<\/jats:italic>\n .\n vivax-<\/jats:italic>\n and\n P<\/jats:italic>\n .\n cynomolgi-<\/jats:italic>\n specific assays with RT were moderately improved at 0.02 and 0.002 parasites\/\u03bcL, respectively (1000-fold change). For DBS\n P<\/jats:italic>\n .\n knowlesi<\/jats:italic>\n samples the use of RT also markedly improved the\n Plasmodium<\/jats:italic>\n genus qPCR LOD from 19.89 to 0.08 parasites\/\u03bcL (249-fold change); no LOD improvement was demonstrated in DBS archived beyond 6 years. The\n Plasmodium<\/jats:italic>\n genus and\n P<\/jats:italic>\n .\n knowlesi<\/jats:italic>\n -assays were 100% specific for\n Plasmodium<\/jats:italic>\n species and\n P<\/jats:italic>\n .\n knowlesi<\/jats:italic>\n detection, respectively, from 190 clinical infections and 48 healthy controls. Reference\n P<\/jats:italic>\n .\n vivax-<\/jats:italic>\n specific primers demonstrated known cross-reactivity with\n P<\/jats:italic>\n .\n cynomolgi<\/jats:italic>\n .\n <\/jats:p>\n <\/jats:sec>\n \n Conclusions\/Significance<\/jats:title>\n \n Our findings support the use of an 18S rRNA\n Plasmodium<\/jats:italic>\n genus qPCR and species-specific nested PCR protocol with RT for highly-sensitive surveillance of zoonotic and human\n Plasmodium<\/jats:italic>\n species infections.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1371\/journal.pntd.0012129","type":"journal-article","created":{"date-parts":[[2025,2,6]],"date-time":"2025-02-06T13:56:31Z","timestamp":1738850191000},"page":"e0012129","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":8,"title":["Improved limit of detection for zoonotic Plasmodium knowlesi and P. cynomolgi surveillance using reverse transcription for total nucleic acid preserved samples or dried blood spots"],"prefix":"10.1371","volume":"19","author":[{"given":"Kamil A.","family":"Braima","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim A.","family":"Piera","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7075-9491","authenticated-orcid":true,"given":"Inke N. D.","family":"Lubis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rintis","family":"Noviyanti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giri S.","family":"Rajahram","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pinkan","family":"Kariodimedjo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irbah R. A.","family":"Nainggolan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ranti","family":"Permatasari","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leily","family":"Trianty","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ristya","family":"Amalia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sitti Saimah","family":"Sakam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angelica F.","family":"Tan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy","family":"William","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacob A. F.","family":"Westaway","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"PingChin","family":"Lee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sylvia","family":"Daim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henry","family":"Surendra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathaniel","family":"Christy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew G.","family":"Letizia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher L.","family":"Peatey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohd Arshil","family":"Moideen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bridget E.","family":"Barber","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Colin J.","family":"Sutherland","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas M.","family":"Anstey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9914-8352","authenticated-orcid":true,"given":"Matthew J.","family":"Grigg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,2,6]]},"reference":[{"issue":"3","key":"pntd.0012129.ref001","doi-asserted-by":"crossref","first-page":"e2780","DOI":"10.1371\/journal.pntd.0002780","article-title":"Defining the Geographical Range of the Plasmodium knowlesi Reservoir.","volume":"8","author":"CL Moyes","year":"2014","journal-title":"PLoS Neglected Trop Dis."},{"key":"pntd.0012129.ref002","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/bs.apar.2021.08.006","article-title":"Epidemiology of the zoonotic malaria Plasmodium knowlesi in changing landscapes.","volume":"113","author":"PR Cuenca","year":"2021","journal-title":"Adv Parasit."},{"issue":"3","key":"pntd.0012129.ref003","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1093\/cid\/cis902","article-title":"A Prospective Comparative Study of Knowlesi, Falciparum, and Vivax Malaria in Sabah, Malaysia: High Proportion With Severe Disease From Plasmodium Knowlesi and Plasmodium Vivax But No Mortality With Early Referral and Artesunate Therapy","volume":"56","author":"BE Barber","year":"2013","journal-title":"Clin Infect Dis"},{"issue":"6","key":"pntd.0012129.ref004","doi-asserted-by":"crossref","first-page":"852","DOI":"10.1086\/605439","article-title":"Clinical and laboratory features of human Plasmodium knowlesi infection","volume":"49","author":"C Daneshvar","year":"2009","journal-title":"Clinical Infectious Diseases"},{"issue":"3","key":"pntd.0012129.ref005","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1093\/cid\/ciy065","article-title":"Age-Related Clinical Spectrum of Plasmodium knowlesi Malaria and Predictors of Severity","volume":"67","author":"MJ Grigg","year":"2018","journal-title":"Clin Infect Dis"},{"issue":"1","key":"pntd.0012129.ref006","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1186\/s12879-021-05941-y","article-title":"Preliminary review on the prevalence, proportion, geographical distribution, and characteristics of naturally acquired Plasmodium cynomolgi infection in mosquitoes, macaques, and humans: a systematic review and meta-analysis.","volume":"21","author":"M Kotepui","year":"2021","journal-title":"BMC Infect Dis"},{"key":"pntd.0012129.ref007","first-page":"77","volume-title":"Advances in Parasitology","author":"MJ Grigg","year":"2021"},{"key":"pntd.0012129.ref008","unstructured":"WHO. Malaria Policy Advisory Group (MPAG) meeting (March 2022) [Internet]. 2022 Apr. Available from: https:\/\/www.who.int\/publications\/i\/item\/9789240048430"},{"issue":"1","key":"pntd.0012129.ref009","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/1475-2875-12-8","article-title":"Limitations of microscopy to differentiate Plasmodium species in a region co-endemic for Plasmodium falciparum, Plasmodium vivax and Plasmodium knowlesi.","volume":"12","author":"BE Barber","year":"2013","journal-title":"Malaria J."},{"issue":"1","key":"pntd.0012129.ref010","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1186\/1475-2875-8-73","article-title":"Morphological features and differential counts of Plasmodium knowlesi parasites in naturally acquired human infections.","volume":"8","author":"KS Lee","year":"2009","journal-title":"Malaria J."},{"issue":"11","key":"pntd.0012129.ref011","doi-asserted-by":"crossref","first-page":"e0006924","DOI":"10.1371\/journal.pntd.0006924","article-title":"Laboratory challenges of Plasmodium species identification in Aceh Province, Indonesia, a malaria elimination setting with newly discovered P. knowlesi.","volume":"12","author":"FN Coutrier","year":"2018","journal-title":"PLoS Neglected Trop Dis"},{"issue":"10","key":"pntd.0012129.ref012","doi-asserted-by":"crossref","first-page":"1936","DOI":"10.3201\/eid2510.190448","article-title":"Plasmodium cynomolgi as Cause of Malaria in Tourist to Southeast Asia, 2018","volume":"25","author":"GN Hartmeyer","year":"2019","journal-title":"Emerging infectious diseases"},{"issue":"1","key":"pntd.0012129.ref013","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1186\/1475-2875-13-68","article-title":"First case of a naturally acquired human infection with Plasmodium cynomolgi.","volume":"13","author":"TH Ta","year":"2014","journal-title":"Malaria J."},{"issue":"4","key":"pntd.0012129.ref014","doi-asserted-by":"crossref","first-page":"1118","DOI":"10.1128\/JCM.03285-12","article-title":"Evaluation of the Sensitivity of a pLDH-Based and an Aldolase-Based Rapid Diagnostic Test for Diagnosis of Uncomplicated and Severe Malaria Caused by PCR-Confirmed Plasmodium knowlesi, Plasmodium falciparum, and Plasmodium vivax","volume":"51","author":"BE Barber","year":"2013","journal-title":"J Clin Microbiol"},{"issue":"6","key":"pntd.0012129.ref015","doi-asserted-by":"crossref","first-page":"2053","DOI":"10.1128\/JCM.00181-14","article-title":"Combining Parasite Lactate Dehydrogenase-Based and Histidine-Rich Protein 2-Based Rapid Tests To Improve Specificity for Diagnosis of Malaria Due to Plasmodium knowlesi and Other Plasmodium Species in Sabah, Malaysia","volume":"52","author":"MJ Grigg","year":"2014","journal-title":"J Clin Microbiol"},{"key":"pntd.0012129.ref016","doi-asserted-by":"crossref","first-page":"1023219","DOI":"10.3389\/fcimb.2022.1023219","article-title":"Diagnostic accuracy and limit of detection of ten malaria parasite lactate dehydrogenase-based rapid tests for Plasmodium knowlesi and P. falciparum.","volume":"12","author":"AF Tan","year":"2022","journal-title":"Front Cell Infect Microbiol"},{"issue":"1","key":"pntd.0012129.ref017","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1186\/s12936-015-0542-5","article-title":"Molecular detection of human Plasmodium species in Sabah using PlasmoNexTM multiplex PCR and hydrolysis probes real-time PCR","volume":"14","author":"PC Lee","year":"2015","journal-title":"Malaria Journal"},{"key":"pntd.0012129.ref018","doi-asserted-by":"crossref","first-page":"39","DOI":"10.12688\/wellcomeopenres.16564.2","article-title":"Reverse transcription PCR to detect low density malaria infections.","volume":"6","author":"P Christensen","year":"2021","journal-title":"Wellcome Open Res"},{"issue":"8","key":"pntd.0012129.ref019","doi-asserted-by":"crossref","first-page":"2946","DOI":"10.1128\/JCM.00276-11","article-title":"Development of a Highly Sensitive Genus-Specific Quantitative Reverse Transcriptase Real-Time PCR Assay for Detection and Quantitation of Plasmodium by Amplifying RNA and DNA of the 18S rRNA Genes","volume":"49","author":"E Kamau","year":"2011","journal-title":"J Clin Microbiol"},{"issue":"11","key":"pntd.0012129.ref020","doi-asserted-by":"crossref","first-page":"1323","DOI":"10.1016\/S0020-7519(02)00111-X","article-title":"Three multigene families in Plasmodium parasites: facts and questions","volume":"32","author":"O Mercereau-Puijalon","year":"2002","journal-title":"Int J Parasitol"},{"issue":"2","key":"pntd.0012129.ref021","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1016\/0166-6851(93)90050-8","article-title":"Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of a high prevalence of mixed infections","volume":"58","author":"G Snounou","year":"1993","journal-title":"Molecular and biochemical parasitology"},{"issue":"4","key":"pntd.0012129.ref022","doi-asserted-by":"crossref","first-page":"e1002015","DOI":"10.1371\/journal.ppat.1002015","article-title":"Plasmodium knowlesi: Reservoir Hosts and Tracking the Emergence in Humans and Macaques.","volume":"7","author":"KS Lee","year":"2011","journal-title":"PLoS Pathog."},{"issue":"5","key":"pntd.0012129.ref023","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1093\/infdis\/jiy520","article-title":"Zoonotic Malaria: The Better You Look, the More You Find","volume":"219","author":"NM Anstey","year":"2019","journal-title":"J Infect Dis"},{"key":"pntd.0012129.ref024","doi-asserted-by":"crossref","first-page":"102589","DOI":"10.1016\/j.parint.2022.102589","article-title":"Improving in vitro continuous cultivation of Plasmodium cynomolgi, a model for P. vivax","volume":"89","author":"P Christensen","year":"2022","journal-title":"Parasitol Int"},{"key":"pntd.0012129.ref025","volume-title":"Microscopy for the detection, identification and quantification of malaria parasites on stained thick and thin blood films in research settings.","author":"WHO","year":"2015"},{"issue":"1","key":"pntd.0012129.ref026","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1186\/s12936-017-2025-3","article-title":"A novel method for extracting nucleic acids from dried blood spots for ultrasensitive detection of low-density Plasmodium falciparum and Plasmodium vivax infections","volume":"16","author":"K Zainabadi","year":"2017","journal-title":"Malar J"},{"issue":"12","key":"pntd.0012129.ref027","doi-asserted-by":"crossref","first-page":"4173","DOI":"10.1128\/JCM.00811-09","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":"M Imwong","year":"2009","journal-title":"J Clin Microbiol"},{"issue":"1","key":"pntd.0012129.ref028","first-page":"344","article-title":"A TaqMan real-time PCR assay for the detection and quantitation of Plasmodium knowlesi. Malar J","volume":"9","author":"PC Divis","year":"2010","journal-title":"Internet]"},{"issue":"7","key":"pntd.0012129.ref029","doi-asserted-by":"crossref","first-page":"1148","DOI":"10.1093\/infdis\/jix091","article-title":"Contribution of Plasmodium knowlesi to Multispecies Human Malaria Infections in North Sumatera, Indonesia","volume":"215","author":"IND Lubis","year":"2017","journal-title":"J Infect Dis"},{"issue":"1","key":"pntd.0012129.ref030","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1186\/s12936-020-03379-2","article-title":"Comparative evaluation of two commercial real-time PCR kits (QuantiFastTM and abTESTM) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia.","volume":"19","author":"NA Nuin","year":"2020","journal-title":"Malaria J."},{"issue":"8","key":"pntd.0012129.ref031","doi-asserted-by":"crossref","first-page":"e0004915","DOI":"10.1371\/journal.pntd.0004915","article-title":"Estimating Geographical Variation in the Risk of Zoonotic Plasmodium knowlesi Infection in Countries Eliminating Malaria. Churcher TS, editor.","volume":"10","author":"FM Shearer","year":"2016","journal-title":"PLoS Neglected Trop Dis [Internet]."},{"issue":"1","key":"pntd.0012129.ref032","doi-asserted-by":"crossref","first-page":"e0011570","DOI":"10.1371\/journal.pntd.0011570","article-title":"Updating estimates of Plasmodium knowlesi malaria risk in response to changing land use patterns across Southeast Asia.","volume":"18","author":"RJ Tobin","year":"2024","journal-title":"PLOS Neglected Trop Dis"},{"issue":"8","key":"pntd.0012129.ref033","first-page":"2187","article-title":"Natural Human Infections with Plasmodium cynomolgi, P. inui, and 4 other Simian Malaria Parasites, Malaysia\u2014Volume 27, Number 8\u2014August 2021\u2014Emerging Infectious Diseases journal\u2014CDC","volume":"27","author":"NJ Yap","year":"2021","journal-title":"Emerg Infect Dis"},{"issue":"5","key":"pntd.0012129.ref034","doi-asserted-by":"crossref","first-page":"784","DOI":"10.1093\/infdis\/jiv475","article-title":"Asymptomatic and Submicroscopic Carriage of Plasmodium knowlesi Malaria in Household and Community Members of Clinical Cases in Sabah, Malaysia","volume":"213","author":"KM Fornace","year":"2016","journal-title":"J Infect Dis"},{"issue":"3","key":"pntd.0012129.ref035","doi-asserted-by":"crossref","first-page":"1107","DOI":"10.4269\/ajtmh.20-0268","article-title":"Prevalence of Asymptomatic and\/or Low-Density Malaria Infection among High-Risk Groups in Peninsular Malaysia","volume":"103","author":"NR Noordin","year":"2020","journal-title":"Am J Trop Med Hyg"},{"issue":"3","key":"pntd.0012129.ref036","doi-asserted-by":"crossref","first-page":"470","DOI":"10.4269\/ajtmh.2007.76.470","article-title":"Submicroscopic Plasmodium falciparum gametocyte densities frequently result in mosquito infection","volume":"76","author":"P Schneider","year":"2007","journal-title":"American Journal of Tropical Medicine and Hygiene"},{"issue":"1","key":"pntd.0012129.ref037","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1186\/s13071-018-2787-7","article-title":"High proportions of asymptomatic and submicroscopic Plasmodium vivax infections in a peri-urban area of low transmission in the Brazilian Amazon.","volume":"11","author":"ACG Almeida","year":"2018","journal-title":"Parasites Vectors"},{"issue":"1","key":"pntd.0012129.ref038","doi-asserted-by":"crossref","first-page":"2945","DOI":"10.1038\/s41467-023-38476-8","article-title":"No evidence of sustained nonzoonotic Plasmodium knowlesi transmission in Malaysia from modelling malaria case data.","volume":"14","author":"KM Fornace","year":"2023","journal-title":"Nat Commun."},{"issue":"9","key":"pntd.0012129.ref039","doi-asserted-by":"crossref","first-page":"3303","DOI":"10.1128\/JCM.01057-14","article-title":"High-throughput ultrasensitive molecular techniques for quantifying low-density malaria parasitemias. Gilligan PH, editor","volume":"52","author":"M Imwong","year":"2014","journal-title":"Journal of clinical microbiology"},{"key":"pntd.0012129.ref040","doi-asserted-by":"crossref","first-page":"105935","DOI":"10.1016\/j.mimet.2020.105935","article-title":"Comparative evaluation of nucleic acid stabilizing reagents for RNA- and DNA-based Leishmania detection in blood as proxy for visceral burdens","volume":"173","author":"E Eberhardt","year":"2020","journal-title":"J Microbiol Methods"},{"issue":"1","key":"pntd.0012129.ref041","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1186\/s12936-015-1038-z","article-title":"An ultrasensitive reverse transcription polymerase chain reaction assay to detect asymptomatic low-density Plasmodium falciparum and Plasmodium vivax infections in small volume blood samples","volume":"14","author":"M Adams","year":"2015","journal-title":"Malar J"},{"issue":"2","key":"pntd.0012129.ref042","doi-asserted-by":"crossref","first-page":"533","DOI":"10.4269\/ajtmh.17-0039","article-title":"A Direct from Blood Reverse Transcriptase Polymerase Chain Reaction Assay for Monitoring Falciparum Malaria Parasite Transmission in Elimination Settings","volume":"97","author":"T Bousema","year":"2017","journal-title":"Am J Trop Med Hyg"},{"issue":"1","key":"pntd.0012129.ref043","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1186\/1475-2875-11-266","article-title":"Filter paper collection of Plasmodium falciparum mRNA for detecting low-density gametocytes.","volume":"11","author":"S Jones","year":"2012","journal-title":"Malar J"},{"issue":"1","key":"pntd.0012129.ref044","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1186\/1475-2875-11-138","article-title":"Stability of gametocyte-specific Pfs25-mRNA in dried blood spots on filter paper subjected to different storage conditions","volume":"11","author":"M Pritsch","year":"2012","journal-title":"Malar J"},{"issue":"2","key":"pntd.0012129.ref045","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1179\/000349803125002977","article-title":"Use of a rapid, single-round, multiplex PCR to detect malarial parasites and identify the species present","volume":"97","author":"D Padley","year":"2003","journal-title":"Ann Trop Med Parasitol"},{"issue":"12","key":"pntd.0012129.ref046","doi-asserted-by":"crossref","first-page":"5636","DOI":"10.1128\/JCM.42.12.5636-5643.2004","article-title":"Detection of Four Plasmodium Species in Blood from Humans by 18S rRNA Gene Subunit-Based and Species-Specific Real-Time PCR Assays","volume":"42","author":"M Rougemont","year":"2004","journal-title":"J Clin Microbiol"},{"issue":"5","key":"pntd.0012129.ref047","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1093\/infdis\/jiy519","article-title":"Asymptomatic Natural Human Infections With the Simian Malaria Parasites Plasmodium cynomolgi and Plasmodium knowlesi","volume":"219","author":"M Imwong","year":"2019","journal-title":"J Infect Dis"},{"issue":"5","key":"pntd.0012129.ref048","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1093\/cid\/ciab1060","article-title":"Cryptic Plasmodium inui and Plasmodium fieldi Infections Among Symptomatic Malaria Patients in Thailand","volume":"75","author":"C Putaporntip","year":"2021","journal-title":"Clin Infect Dis"},{"issue":"7","key":"pntd.0012129.ref049","first-page":"1232","article-title":"Co-infections of Plasmodium knowlesi, P. falciparum, and P. vivax among Humans and Anopheles dirus Mosquitoes, Southern Vietnam\u2014Volume 17, Number 7\u2014July 2011\u2014Emerging Infectious Diseases journal\u2014CDC","volume":"17","author":"RP Marchand","year":"2011","journal-title":"Emerg Infect Dis"}],"updated-by":[{"DOI":"10.1371\/journal.pntd.0012129","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2025,2,19]],"date-time":"2025-02-19T00:00:00Z","timestamp":1739923200000}}],"container-title":["PLOS Neglected Tropical Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pntd.0012129","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,2,19]],"date-time":"2025-02-19T13:39:45Z","timestamp":1739972385000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pntd.0012129"}},"subtitle":[],"editor":[{"given":"Richard","family":"Culleton","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2025,2,6]]},"references-count":49,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2025,2,6]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pntd.0012129","relation":{},"ISSN":["1935-2735"],"issn-type":[{"value":"1935-2735","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,6]]}}}