{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T18:25:34Z","timestamp":1783535134422,"version":"3.55.0"},"reference-count":41,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,1,2]],"date-time":"2026-01-02T00:00:00Z","timestamp":1767312000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      Malaria is still a public health challenge across many regions of the world. With significant efforts, it has become a target for elimination in near future. However, elimination requires early and accurate diagnosis and despite the fact that malaria elimination is nearing in several countries, specific and sensitive diagnosis of malaria and its causative species remains inadequate in many near-elimination settings, including India. With the advent and increasing usage of nucleic acid-based detection of\n                      <jats:italic>Plasmodium<\/jats:italic>\n                      infection, the diagnostic limitations of microscopy, including errors in species identification, have become more prominent.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      The purpose of this study was to determine the magnitude of\n                      <jats:italic>Plasmodium<\/jats:italic>\n                      species misidentification by microscopy in India based on previously published reports that performed microscopy and PCR to the same samples. A total of 2706 microscopy-PCR pairs were extracted from 16 different locations across 11 Indian states. Region-specific and species-wise misidentification rates were also estimated.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The analyses revealed 15% misidentification rate (408\/2706). Surprisingly, microscopy misidentified &amp;gt;98% of mixed-infections (400\/405) as mono-infections (almost all as\n                      <jats:italic>P. falciparum<\/jats:italic>\n                      mono-infections). The study identifies Jharkhand and Madhya Pradesh as major contributors (&amp;gt;20%) to\n                      <jats:italic>Plasmodium<\/jats:italic>\n                      species misidentification by microscopy.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      These findings suggest that we are overestimating\n                      <jats:italic>P. falciparum<\/jats:italic>\n                      burden, potentially wasting elimination resources, and underestimating non-falciparum species. The study also addresses an important issue concerning analysis of misidentification and sub-microscopic infection data. It proposes an analysis approach that is expected to help in deciphering misidentification and sub-microscopic infections in a more granular manner, generating actionable data for countries targeting malaria elimination.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcimb.2025.1695062","type":"journal-article","created":{"date-parts":[[2026,1,2]],"date-time":"2026-01-02T06:28:28Z","timestamp":1767335308000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Plasmodium mono and mixed-infections in India: a tale of see-saw in species misidentification"],"prefix":"10.3389","volume":"15","author":[{"given":"Nimita","family":"Deora","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Veena","family":"Pande","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abhinav","family":"Sinha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,1,2]]},"reference":[{"key":"B1","unstructured":"Worldwide E-Learning Course on Malaria Microscopy (WELCOMM)"},{"key":"B2","volume-title":"World malaria report 2024: addressing inequity in the global malaria response","year":"2024"},{"key":"B3","volume-title":"WHO guidelines for malaria, 16 October 2023","year":"2023"},{"key":"B4","volume-title":"Malaria surveillance, monitoring & evaluation: a reference manual","year":"2018"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.3389\/frai.2022.510483","article-title":"Digital microscopy and artificial intelligence could profoundly contribute to malaria diagnosis in elimination settings","volume":"5","author":"Beck","year":"2022","journal-title":"Front. Artif. Intell."},{"key":"B6","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1111\/tmi.12005","article-title":"Emergence of a new focus of Plasmodium malariae in forest villages of district Balaghat, Central India: implications for the diagnosis of malaria and its control","volume":"18","author":"Bharti","year":"2013","journal-title":"Trop. Med. Int. Health"},{"key":"B7","doi-asserted-by":"publisher","first-page":"2265","DOI":"10.3390\/microorganisms9112265","article-title":"Malaria diagnosis in non-endemic settings: the European experience in the last 22 years","volume":"9","author":"Calderaro","year":"2021","journal-title":"Microorganismicroscopy."},{"key":"B8","doi-asserted-by":"publisher","first-page":"352","DOI":"10.1093\/trstmh\/trv015","article-title":"Sympatric distribution of Plasmodium ovale curtisi and P. ovale wallikeri in India: implication for the diagnosis of malaria and its control","volume":"109","author":"Chaturvedi","year":"2015","journal-title":"Trans. R Soc. Trop. Med. Hyg."},{"key":"B9","doi-asserted-by":"publisher","first-page":"100190","DOI":"10.1016\/j.onehlt.2020.100190","article-title":"Trends of neglected Plasmodium species infection in humans over the past century in India","volume":"11","author":"Chaturvedi","year":"2020","journal-title":"One Health"},{"key":"B10","doi-asserted-by":"publisher","first-page":"935","DOI":"10.1016\/j.pt.2021.07.017","article-title":"Plasmodium and malaria: adding to the don\u2019ts","volume":"37","author":"Chauhan","year":"2021","journal-title":"Trends Parasitol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"1259","DOI":"10.3390\/pathogens11070737","article-title":"Multiplexing for Plasmodium spp.? Think Again! Comment on Bhowmick et al. Dry Post Wintertime Mass Surveillance Unearths a Huge Burden of P. vivax, and Mixed Infection with P. vivax P. falciparum, a Threat to Malaria Elimination, in Dhalai, Tripura, India","volume":"10","author":"Deora","year":"2021","journal-title":"Pathogens"},{"key":"B12","doi-asserted-by":"publisher","first-page":"e019170","DOI":"10.1136\/bmjgh-2025-019170","article-title":"Time to address the drawbacks in estimation and reporting of malaria cases in India and beyond","volume":"10","author":"Deora","year":"2025","journal-title":"BMJ Glob Health"},{"key":"B13","doi-asserted-by":"publisher","first-page":"100056","DOI":"10.1016\/j.lansea.2022.100056","article-title":"Truenat\u00ae for Plasmodium sub-microscopic infections: Miles to go\u2026","volume":"5","author":"Deora","year":"2022","journal-title":"Lancet Reg. Health Southeast Asia."},{"key":"B14","doi-asserted-by":"publisher","first-page":"100012","DOI":"10.1016\/j.lansea.2022.05.001","article-title":"A systematic review and meta-analysis on sub-microscopic Plasmodium infections in India: Different perspectives and global challenges","volume":"2","author":"Deora","year":"2022","journal-title":"Lancet Regional Health - Southeast Asia."},{"key":"B15","doi-asserted-by":"publisher","first-page":"401","DOI":"10.14411\/fp.2013.041","article-title":"Nested PCR detection of Plasmodium malariae from microscopy confirmed P. falciparum samples in endemic area of NE India","volume":"60","author":"Dhiman","year":"2013","journal-title":"Folia Parasitol. (Praha)."},{"key":"B16","doi-asserted-by":"publisher","first-page":"719","DOI":"10.1309\/LM0KC4BEYHGDSZCU","article-title":"Detection and identification of malaria parasites: A review of proficiency test results and laboratory practices","volume":"41","author":"Edson","year":"2010","journal-title":"Lab. Med."},{"key":"B17","doi-asserted-by":"publisher","first-page":"S4","DOI":"10.1016\/S2214-109X(15)70123-3","article-title":"Online training and educational games for malaria diagnosis","volume":"3","author":"Feng","year":"2015","journal-title":"Lancet Global Health"},{"key":"B18","doi-asserted-by":"publisher","first-page":"21807","DOI":"10.1038\/s41598-020-78691-7","article-title":"Misidentification of Plasmodium ovale as Plasmodium vivax malaria by a microscopic method: a meta-analysis of confirmed P. ovale cases","volume":"10","author":"Kotepui","year":"2020","journal-title":"Sci. Rep."},{"key":"B19","doi-asserted-by":"publisher","first-page":"1853","DOI":"10.3201\/eid2110.150678","article-title":"Detection of mixed infections with plasmodium spp. by PCR, India, 2014","volume":"21","author":"Krishna","year":"2015","journal-title":"Emerg. Infect. Dis."},{"key":"B20","doi-asserted-by":"publisher","first-page":"16860","DOI":"10.1038\/s41598-017-16974-2","article-title":"Prevalence of malaria in two highly endemic Community Health Centers in the Bastar district, Chhattisgarh showing mixed infections with Plasmodium species","volume":"7","author":"Krishna","year":"2017","journal-title":"Sci. Rep."},{"key":"B21","doi-asserted-by":"publisher","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":"Lee","year":"2009","journal-title":"Malar J."},{"key":"B22","doi-asserted-by":"publisher","first-page":"92","DOI":"10.1186\/1475-2875-5-92","article-title":"Production and validation of durable, high quality standardized malaria microscopy slides for teaching, testing and quality assurance during an era of declining diagnostic proficiency","volume":"5","author":"Maguire","year":"2006","journal-title":"Malar J."},{"key":"B23","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1186\/s12936-021-03714-1","article-title":"Quantification of the misidentification of Plasmodium knowlesi as Plasmodium malariae by microscopy: an analysis of 1569 P","volume":"20","author":"Mahittikorn","year":"2021","journal-title":"knowlesi cases. Malar J."},{"key":"B24","doi-asserted-by":"publisher","first-page":"2018:4914358","DOI":"10.1155\/2018\/4914358","article-title":"Evaluation of routine microscopy performance for malaria diagnosis at three different health centers in Brazzaville, Republic of Congo","author":"Mayengue","year":"2018","journal-title":"Malar Res. Treat."},{"key":"B25","doi-asserted-by":"publisher","first-page":"372","DOI":"10.4269\/ajtmh.2003.69.372","article-title":"Dependence of malaria detection and species diagnosis by microscopy on parasite density","volume":"69","author":"McKenzie","year":"2003","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"B26","doi-asserted-by":"publisher","first-page":"52","DOI":"10.25259\/IJMR_20081281_52","article-title":"Detection & molecular confirmation of a focus of Plasmodium malariae in Arunachal Pradesh, India","volume":"128","author":"Mohapatra","year":"2008","journal-title":"Indian J. Med. Res."},{"key":"B27","doi-asserted-by":"publisher","first-page":"437","DOI":"10.1186\/1475-2875-13-437","article-title":"Impact of a training course on the quality of malaria diagnosis by microscopy in Angola","volume":"13","author":"Moura","year":"2014","journal-title":"Malar J."},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0146450","article-title":"Performance of microscopy for the diagnosis of malaria and human African trypanosomiasis by diagnostic laboratories in the democratic republic of the congo: results of a nation-wide external quality assessment","volume":"11","author":"Mukadi","year":"2016","journal-title":"PloS One"},{"key":"B29","volume-title":"Malaria Elimination in India","year":"2017"},{"key":"B30","volume-title":"Laboratory diagnosis of malaria. Operational guidelines for laboratory technicians","year":"2007"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1016\/j.lansea.2022.100054","article-title":"Strengthening malaria microscopy using artificial intelligence-based approaches in India","volume":"5","author":"Nema","year":"2022","journal-title":"Lancet Reg. Health Southeast Asia."},{"key":"B32","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1186\/1475-2875-12-113","article-title":"Misclassification of Plasmodium infections by conventional microscopy and the impact of remedial training on the proficiency of laboratory technicians in species identification","volume":"12","author":"Obare","year":"2013","journal-title":"Malar J."},{"key":"B33","doi-asserted-by":"publisher","first-page":"1237","DOI":"10.1038\/ncomms2241","article-title":"Factors determining the occurrence of submicroscopic malaria infections and their relevance for control","volume":"3","author":"Okell","year":"2012","journal-title":"Nat. Commun."},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0008410","article-title":"Substantial contribution of submicroscopical Plasmodium falciparum gametocyte carriage to the infectious reservoir in an area of seasonal transmission","volume":"4","author":"Ou\u00e9draogo","year":"2009","journal-title":"PloS One"},{"key":"B35","doi-asserted-by":"publisher","first-page":"233ed9","DOI":"10.1126\/scitranslmed.3009172","article-title":"Educational games for malaria diagnosis","volume":"6","author":"Ozcan","year":"2014","journal-title":"Sci. Transl. Med."},{"key":"B36","doi-asserted-by":"publisher","first-page":"310","DOI":"10.1186\/s12936-015-0824-y","article-title":"Therapeutic assessment of chloroquine-primaquine combined regimen in adult cohort of Plasmodium vivax malaria from a tertiary care hospital in southwestern India","volume":"14","author":"Rishikesh","year":"2015","journal-title":"Malar J."},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0157666","article-title":"Therapeutic assessment of chloroquine-primaquine combined regimen in adult cohort of plasmodium vivax malaria from primary care centres in Southwestern India","volume":"11","author":"Saravu","year":"2016","journal-title":"PloS One"},{"key":"B38","doi-asserted-by":"publisher","first-page":"1416","DOI":"10.1111\/tmi.12184","article-title":"First report of detection and molecular confirmation of Plasmodium ovale from severe malaria cases in central India","volume":"18","author":"Singh","year":"2013","journal-title":"Trop. Med. Int. Health"},{"key":"B39","volume-title":"Malaria microscopy quality assurance manual, version2","year":"2016"},{"key":"B40","article-title":"Meeting report of the WHO evidence review group on low-density malaria infections","year":"2017"},{"key":"B41","article-title":"Technical consultation on research requirements to support policy recommendations on highly sensitive point-of-care diagnostics for P. falciparum malaria","year":"2018"}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2025.1695062\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,2]],"date-time":"2026-01-02T06:28:29Z","timestamp":1767335309000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2025.1695062\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,2]]},"references-count":41,"alternative-id":["10.3389\/fcimb.2025.1695062"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2025.1695062","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,2]]},"article-number":"1695062"}}