{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,21]],"date-time":"2026-06-21T06:13:46Z","timestamp":1782022426307,"version":"3.54.5"},"reference-count":29,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,1,2]],"date-time":"2021-01-02T00:00:00Z","timestamp":1609545600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2021,1,2]],"date-time":"2021-01-02T00:00:00Z","timestamp":1609545600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000009","name":"Foundation for the National Institutes of Health","doi-asserted-by":"publisher","award":["AI089674"],"award-info":[{"award-number":["AI089674"]}],"id":[{"id":"10.13039\/100000009","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000061","name":"Fogarty International Center","doi-asserted-by":"crossref","award":["TW010526"],"award-info":[{"award-number":["TW010526"]}],"id":[{"id":"10.13039\/100000061","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100005473","name":"Belfer Center for Science and International Affairs, Harvard University","doi-asserted-by":"crossref","award":["TW010365"],"award-info":[{"award-number":["TW010365"]}],"id":[{"id":"10.13039\/100005473","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Malar J"],"published-print":{"date-parts":[[2021,12]]},"abstract":"Abstract<\/jats:title>\n \n Background<\/jats:title>\n \n Rapid diagnostic tests (RDTs) play a key role in malaria case management. The most widely used RDT identifies\n Plasmodium falciparum<\/jats:italic>\n based on immunochromatographic recognition of\n P. falciparum<\/jats:italic>\n histidine-rich protein 2 (PfHRP2). Deletion of the paralogous\n pfhrp2<\/jats:italic>\n and\n pfhrp3<\/jats:italic>\n genes leads to false-negative PfHRP2-based RDTs, and has been reported in\n P. falciparum<\/jats:italic>\n infections from South America and Africa. However, identification of\n pfhrp2\/pfhrp3<\/jats:italic>\n deletions has usually been based only on failure to amplify these genes using PCR, without confirmation based on PfHRP2 protein expression, and understanding of the true prevalence of deletions is incomplete.\n <\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n Deletions of\n pfhrp2<\/jats:italic>\n \/\n pfhrp3<\/jats:italic>\n in blood samples were investigated from cross-sectional surveys in 2012-13 in three regions of varied malaria transmission intensity in Uganda. Samples with positive Giemsa-stained thick blood smears, but negative PfHRP2-based RDTs were evaluated by PCR amplification of conserved subunit ribosomal DNA for\n Plasmodium<\/jats:italic>\n species, PCR amplification of\n pfhrp2<\/jats:italic>\n and\n pfhrp3<\/jats:italic>\n genes to identify deletions, and bead-based immunoassays for expression of PfHRP2.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n Of 3516 samples collected in cross-sectional surveys, 1493 (42.5%) had positive blood smears, of which 96 (6.4%) were RDT-negative. Of these 96 RDT-negative samples,\n P. falciparum<\/jats:italic>\n DNA was identified by PCR in 56 (58%) and only non-falciparum plasmodial DNA in 40 (42%). In all 56\n P. falciparum<\/jats:italic>\n -positive samples there was a failure to amplify\n pfhrp2<\/jats:italic>\n or\n pfhrp3<\/jats:italic>\n : in 25 (45%)\n pfhrp2<\/jats:italic>\n was not amplified, in 39 (70%)\n pfhrp3<\/jats:italic>\n was not amplified, and in 19 (34%) neither gene was amplified. For the 39\n P. falciparum<\/jats:italic>\n -positive, RDT-negative samples available for analysis of protein expression, PfHRP2 was not identified by immunoassay in only four samples (10.3%); these four samples all had failure to amplify both\n pfhrp2<\/jats:italic>\n and\n pfhrp3<\/jats:italic>\n by PCR. Thus, only four of 96 (4.2%) smear-positive, RDT-negative samples had\n P. falciparum<\/jats:italic>\n infections with deletion of\n pfhrp2<\/jats:italic>\n and\n pfhrp3<\/jats:italic>\n confirmed by failure to amplify the genes by PCR and lack of expression of PfHRP2 demonstrated by immunoassay.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion<\/jats:title>\n \n False negative RDTs were uncommon. Deletions in\n pfhrp2<\/jats:italic>\n and\n pfhrp3<\/jats:italic>\n explained some of these false negatives, but most false negatives were not due to deletion of the\n pfhrp2<\/jats:italic>\n and\n pfhrp3<\/jats:italic>\n genes.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1186\/s12936-020-03547-4","type":"journal-article","created":{"date-parts":[[2021,1,1]],"date-time":"2021-01-01T23:17:11Z","timestamp":1609543031000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Deletions of pfhrp2 and pfhrp3 genes were uncommon in rapid diagnostic test-negative Plasmodium falciparum isolates from Uganda"],"prefix":"10.1186","volume":"20","author":[{"given":"Sam L.","family":"Nsobya","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew","family":"Walakira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth","family":"Namirembe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moses","family":"Kiggundu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joaniter I.","family":"Nankabirwa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emmanuel","family":"Ruhamyankaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emmanuel","family":"Arinaitwe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melissa D.","family":"Conrad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moses R.","family":"Kamya","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grant","family":"Dorsey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip J.","family":"Rosenthal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,1,2]]},"reference":[{"key":"3547_CR1","unstructured":"WHO. World malaria report. Geneva, Global Malaria Programme, World Health Organization, 2019:232."},{"key":"3547_CR2","doi-asserted-by":"publisher","first-page":"112","DOI":"10.1016\/j.pt.2019.12.004","volume":"36","author":"KE Poti","year":"2020","unstructured":"Poti KE, Sullivan DJ, Dondorp AM, Woodrow CJ. HRP2: transforming malaria diagnosis, but with caveats. Trends Parasitol. 2020;36:112\u201326.","journal-title":"Trends Parasitol."},{"key":"3547_CR3","doi-asserted-by":"publisher","first-page":"331","DOI":"10.1016\/j.trstmh.2006.09.007","volume":"101","author":"PL Chiodini","year":"2007","unstructured":"Chiodini PL, Bowers K, Jorgensen P, Barnwell JW, Grady KK, Luchavez J, et al. The heat stability of Plasmodium lactate dehydrogenase-based and histidine-rich protein 2-based malaria rapid diagnostic tests. Trans R Soc Trop Med Hyg. 2007;101:331\u20137.","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"3547_CR4","doi-asserted-by":"publisher","first-page":"2773","DOI":"10.1128\/JCM.02557-05","volume":"44","author":"N Lee","year":"2006","unstructured":"Lee N, Baker J, Andrews KT, Gatton ML, Bell D, Cheng Q, et al. Effect of sequence variation in Plasmodium falciparum histidine- rich protein 2 on binding of specific monoclonal antibodies: implications for rapid diagnostic tests for malaria. J Clin Microbiol. 2006;44:2773\u20138.","journal-title":"J Clin Microbiol"},{"key":"3547_CR5","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1186\/s12936-014-0537-7","volume":"14","author":"JF Abdallah","year":"2015","unstructured":"Abdallah JF, Okoth SA, Fontecha GA, Torres RE, Banegas EI, Matute ML, et al. Prevalence of pfhrp2 and pfhrp3 gene deletions in Puerto Lempira, Honduras. Malar J. 2015;14:19.","journal-title":"Malar J."},{"key":"3547_CR6","doi-asserted-by":"publisher","first-page":"e25008","DOI":"10.7554\/eLife.25008","volume":"6","author":"OJ Watson","year":"2017","unstructured":"Watson OJ, Slater HC, Verity R, Parr JB, Mwandagalirwa MK, Tshefu A, et al. Modelling the drivers of the spread of Plasmodium falciparum hrp2 gene deletions in sub-Saharan Africa. Elife. 2017;6:e25008.","journal-title":"Elife."},{"key":"3547_CR7","doi-asserted-by":"publisher","first-page":"1156","DOI":"10.1093\/infdis\/jix094","volume":"215","author":"ML Gatton","year":"2017","unstructured":"Gatton ML, Dunn J, Chaudhry A, Ciketic S, Cunningham J, Cheng Q. Implications of parasites lacking Plasmodium falciparum histidine-rich protein 2 on malaria morbidity and control when rapid diagnostic tests are used for diagnosis. J Infect Dis. 2017;215:1156\u201366.","journal-title":"J Infect Dis"},{"key":"3547_CR8","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1007\/s11908-018-0655-4","volume":"20","author":"AN Mukkala","year":"2018","unstructured":"Mukkala AN, Kwan J, Lau R, Harris D, Kain D, Boggild AK. An update on malaria rapid diagnostic tests. Curr Infect Dis Rep. 2018;20:49.","journal-title":"Curr Infect Dis Rep."},{"key":"3547_CR9","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1186\/1475-2875-12-34","volume":"12","author":"N Wurtz","year":"2013","unstructured":"Wurtz N, Fall B, Bui K, Pascual A, Fall M, Camara C, et al. Pfhrp2 and pfhrp3 polymorphisms in Plasmodium falciparum isolates from Dakar, Senegal: impact on rapid malaria diagnostic tests. Malar J. 2013;12:34.","journal-title":"Malar J."},{"key":"3547_CR10","doi-asserted-by":"publisher","first-page":"2694","DOI":"10.1128\/JCM.00281-11","volume":"49","author":"S Houze","year":"2011","unstructured":"Houze S, Hubert V, Le Pessec G, Le Bras J, Clain J. Combined deletions of pfhrp2 and pfhrp3 genes result in Plasmodium falciparum malaria false-negative rapid diagnostic test. J Clin Microbiol. 2011;49:2694\u20136.","journal-title":"J Clin Microbiol"},{"key":"3547_CR11","doi-asserted-by":"publisher","first-page":"160","DOI":"10.1186\/1475-2875-7-160","volume":"7","author":"SA Harvey","year":"2008","unstructured":"Harvey SA, Jennings L, Chinyama M, Masaninga F, Mulholland K, Bell DR. Improving community health worker use of malaria rapid diagnostic tests in Zambia: package instructions, job aid and job aid-plus-training. Malar J. 2008;7:160.","journal-title":"Malar J."},{"key":"3547_CR12","doi-asserted-by":"crossref","unstructured":"Gamboa D, Ho MF, Bendezu J, Torres K, Chiodini PL, Barnwell JW, et al. A large proportion of P. falciparum isolates in the Amazon region of Peru lack pfhrp2 and pfhrp3: implications for malaria rapid diagnostic tests. PLoS One. 2010;5:e8091.","DOI":"10.1371\/journal.pone.0008091"},{"key":"3547_CR13","doi-asserted-by":"publisher","first-page":"462","DOI":"10.3201\/eid2403.171723","volume":"24","author":"A Berhane","year":"2018","unstructured":"Berhane A, Anderson K, Mihreteab S, Gresty K, Rogier E, Mohamed S, et al. Major threat to malaria control programs by Plasmodium falciparum lacking histidine-rich protein 2, Eritrea. Emerg Infect Dis. 2018;24:462\u201370.","journal-title":"Emerg Infect Dis."},{"key":"3547_CR14","doi-asserted-by":"publisher","first-page":"194","DOI":"10.4269\/ajtmh.2012.10-0665","volume":"86","author":"OA Koita","year":"2012","unstructured":"Koita OA, Doumbo OK, Ouattara A, Tall LK, Konare A, Diakite M, et al. False-negative rapid diagnostic tests for malaria and deletion of the histidine-rich repeat region of the hrp2 gene. Am J Trop Med Hyg. 2012;86:194\u20138.","journal-title":"Am J Trop Med Hyg"},{"key":"3547_CR15","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1186\/s12936-017-1768-1","volume":"16","author":"CT Kozycki","year":"2017","unstructured":"Kozycki CT, Umulisa N, Rulisa S, Mwikarago EI, Musabyimana JP, Habimana JP, et al. False-negative malaria rapid diagnostic tests in Rwanda: impact of Plasmodium falciparum isolates lacking hrp2 and declining malaria transmission. Malar J. 2017;16:123.","journal-title":"Malar J."},{"key":"3547_CR16","doi-asserted-by":"publisher","first-page":"36","DOI":"10.1093\/infdis\/jix347","volume":"216","author":"JB Parr","year":"2017","unstructured":"Parr JB, Verity R, Doctor SM, Janko M, Carey-Ewend K, Turman BJ, et al. Pfhrp2-Deleted Plasmodium falciparum parasites in the Democratic Republic of the Congo: a national cross-sectional survey. J Infect Dis. 2017;216:36\u201344.","journal-title":"J Infect Dis"},{"key":"3547_CR17","doi-asserted-by":"publisher","first-page":"14718","DOI":"10.1038\/s41598-017-15031-2","volume":"7","author":"KB Beshir","year":"2017","unstructured":"Beshir KB, Sepulveda N, Bharmal J, Robinson A, Mwanguzi J, Busula AO, et al. Plasmodium falciparum parasites with histidine-rich protein 2 (pfhrp2) and pfhrp3 gene deletions in two endemic regions of Kenya. Sci Rep. 2017;7:14718.","journal-title":"Sci Rep."},{"key":"3547_CR18","doi-asserted-by":"publisher","first-page":"1444","DOI":"10.1093\/infdis\/jiz335","volume":"220","author":"R Thomson","year":"2019","unstructured":"Thomson R, Beshir KB, Cunningham J, Baiden F, Bharmal J, Bruxvoort KJ, et al. pfhrp2 and pfhrp3 gene deletions that affect malaria rapid diagnostic tests for Plasmodium falciparum: analysis of archived blood samples from 3 African countries. J Infect Dis. 2019;220:1444\u201352.","journal-title":"J Infect Dis"},{"key":"3547_CR19","doi-asserted-by":"publisher","first-page":"300","DOI":"10.1186\/s12936-020-03362-x","volume":"19","author":"AB Bosco","year":"2020","unstructured":"Bosco AB, Anderson K, Gresty K, Prosser C, Smith D, Nankabirwa JI, et al. Molecular surveillance reveals the presence of pfhrp2 and pfhrp3 gene deletions in Plasmodium falciparum parasite populations in Uganda, 2017-2019. Malar J. 2020;19:300.","journal-title":"Malar J."},{"key":"3547_CR20","doi-asserted-by":"publisher","first-page":"e1002167","DOI":"10.1371\/journal.pmed.1002167","volume":"13","author":"A Katureebe","year":"2016","unstructured":"Katureebe A, Zinszer K, Arinaitwe E, Rek J, Kakande E, Charland K, et al. Measures of malaria burden after long-lasting insecticidal net distribution and indoor residual spraying at three sites in Uganda: a prospective observational study. PLoS Med. 2016;13:e1002167.","journal-title":"PLoS Med."},{"key":"3547_CR21","doi-asserted-by":"publisher","first-page":"528","DOI":"10.1186\/s12936-015-1056-x","volume":"14","author":"JI Nankabirwa","year":"2015","unstructured":"Nankabirwa JI, Yeka A, Arinaitwe E, Kigozi R, Drakeley C, Kamya MR, et al. Estimating malaria parasite prevalence from community surveys in Uganda: a comparison of microscopy, rapid diagnostic tests and polymerase chain reaction. Malar J. 2015;14:528.","journal-title":"Malar J."},{"key":"3547_CR22","doi-asserted-by":"publisher","first-page":"528","DOI":"10.1128\/JCM.43.1.528.2005","volume":"43","author":"JC Petithory","year":"2005","unstructured":"Petithory JC, Ardoin F, Ash LR. Rapid and inexpensive method of diluting Giemsa stain for diagnosis of malaria and other infestations by blood parasites. J Clin Microbiol. 2005;43:528.","journal-title":"J Clin Microbiol"},{"key":"3547_CR23","doi-asserted-by":"publisher","first-page":"565","DOI":"10.4269\/ajtmh.1995.52.565","volume":"52","author":"CV Plowe","year":"1995","unstructured":"Plowe CV, Djimde A, Bouare M, Doumbo O, Wellems TE. Pyrimethamine and proguanil resistance-conferring mutations in Plasmodium falciparum dihydrofolate reductase: polymerase chain reaction methods for surveillance in Africa. Am J Trop Med Hyg. 1995;52:565\u20138.","journal-title":"Am J Trop Med Hyg"},{"key":"3547_CR24","doi-asserted-by":"publisher","first-page":"283","DOI":"10.1016\/0166-6851(93)90050-8","volume":"58","author":"G Snounou","year":"1993","unstructured":"Snounou G, Viriyakosol S, Jarra W, Thaithong S, Brown KN. 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. Mol Biochem Parasitol. 1993;58:283\u201392.","journal-title":"Mol Biochem Parasitol"},{"key":"3547_CR25","doi-asserted-by":"publisher","first-page":"133","DOI":"10.4269\/ajtmh.2003.68.133","volume":"68","author":"A Cattamanchi","year":"2003","unstructured":"Cattamanchi A, Kyabayinze D, Hubbard A, Rosenthal PJ, Dorsey G. Distinguishing recrudescence from reinfection in a longitudinal antimalarial drug efficacy study: comparison of results based on genotyping of msp-1, msp-2, and glurp. Am J Trop Med Hyg. 2003;68:133\u20139.","journal-title":"Am J Trop Med Hyg"},{"key":"3547_CR26","doi-asserted-by":"publisher","first-page":"685","DOI":"10.1093\/infdis\/jix622","volume":"217","author":"MM Plucinski","year":"2018","unstructured":"Plucinski MM, Dimbu PR, Fortes F, Abdulla S, Ahmed S, Gutman J, et al. Posttreatment HRP2 clearance in patients with uncomplicated Plasmodium falciparum malaria. J Infect Dis. 2018;217:685\u201392.","journal-title":"J Infect Dis"},{"key":"3547_CR27","doi-asserted-by":"publisher","first-page":"402","DOI":"10.1186\/s12936-019-3027-0","volume":"18","author":"E Rogier","year":"2019","unstructured":"Rogier E, van den Hoogen L, Herman C, Gurrala K, Joseph V, Stresman G, et al. High-throughput malaria serosurveillance using a one-step multiplex bead assay. Malar J. 2019;18:402.","journal-title":"Malar J."},{"key":"3547_CR28","doi-asserted-by":"publisher","first-page":"e0172139","DOI":"10.1371\/journal.pone.0172139","volume":"12","author":"E Rogier","year":"2017","unstructured":"Rogier E, Plucinski M, Lucchi N, Mace K, Chang M, Lemoine JF, et al. Bead-based immunoassay allows sub-picogram detection of histidine-rich protein 2 from Plasmodium falciparum and estimates reliability of malaria rapid diagnostic tests. PLoS ONE. 2017;12:e0172139.","journal-title":"PLoS ONE"},{"key":"3547_CR29","doi-asserted-by":"publisher","first-page":"753","DOI":"10.4269\/ajtmh.17-0345","volume":"97","author":"V Asua","year":"2017","unstructured":"Asua V, Tukwasibwe S, Conrad M, Walakira A, Nankabirwa JI, Mugenyi L, et al. Plasmodium species infecting children presenting with malaria in Uganda. Am J Trop Med Hyg. 2017;97:753\u20137.","journal-title":"Am J Trop Med Hyg"}],"container-title":["Malaria Journal"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/link.springer.com\/content\/pdf\/10.1186\/s12936-020-03547-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/link.springer.com\/article\/10.1186\/s12936-020-03547-4\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/link.springer.com\/content\/pdf\/10.1186\/s12936-020-03547-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,1,1]],"date-time":"2021-01-01T23:17:28Z","timestamp":1609543048000},"score":1,"resource":{"primary":{"URL":"https:\/\/malariajournal.biomedcentral.com\/articles\/10.1186\/s12936-020-03547-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1,2]]},"references-count":29,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2021,12]]}},"alternative-id":["3547"],"URL":"https:\/\/doi.org\/10.1186\/s12936-020-03547-4","relation":{"has-preprint":[{"id-type":"doi","id":"10.21203\/rs.3.rs-42855\/v1","asserted-by":"object"},{"id-type":"doi","id":"10.21203\/rs.3.rs-42855\/v2","asserted-by":"object"}]},"ISSN":["1475-2875"],"issn-type":[{"value":"1475-2875","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,1,2]]},"assertion":[{"value":"20 July 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"12 December 2020","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"2 January 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"All authors declare that they have no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"4"}}