{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,5]],"date-time":"2026-11-05T19:39:11Z","timestamp":1793907551387,"version":"4.8.0"},"reference-count":37,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2020,10,9]],"date-time":"2020-10-09T00:00:00Z","timestamp":1602201600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2020,10,9]],"date-time":"2020-10-09T00:00:00Z","timestamp":1602201600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100001349","name":"Singapore National Medical Research Council","doi-asserted-by":"crossref","award":["NMRC\/OFIRG\/0040\/2017"],"award-info":[{"award-number":["NMRC\/OFIRG\/0040\/2017"]}],"id":[{"id":"10.13039\/501100001349","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001459","name":"Singapore Ministry of Education","doi-asserted-by":"crossref","award":["MOE2017-T2-2-030 (S) AcRF Tier 2"],"award-info":[{"award-number":["MOE2017-T2-2-030 (S) AcRF Tier 2"]}],"id":[{"id":"10.13039\/501100001459","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Malar J"],"published-print":{"date-parts":[[2020,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Sequencing technology advancements opened new opportunities to use transcriptomics for studying malaria pathology and epidemiology. Even though in recent years the study of whole parasite transcriptome proved to be essential in understanding parasite biology there is no compiled up-to-date reference protocol for the efficient generation of transcriptome data from growing number of samples. Here, a comprehensive methodology on how to preserve, extract, amplify, and sequence full-length mRNA transcripts from\n                      <jats:italic>Plasmodium<\/jats:italic>\n                      -infected blood samples is presented that can be fully streamlined for high-throughput studies.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The utility of various commercially available RNA-preserving reagents in a range of storage conditions was evaluated. Similarly, several RNA extraction protocols were compared and the one most suitable method for the extraction of high-quality total RNA from low-parasitaemia and low-volume blood samples was established. Furthermore, the criteria needed to evaluate the quality and integrity of\n                      <jats:italic>Plasmodium<\/jats:italic>\n                      RNA in the presence of human RNA was updated. Optimization of SMART-seq2 amplification method to better suit AT-rich\n                      <jats:italic>Plasmodium falciparum<\/jats:italic>\n                      RNA samples allowed us to generate high-quality transcriptomes from as little as 10\u00a0ng of total RNA and a lower parasitaemia limit of 0.05%. Finally, a modified method for depletion of unwanted human haemoglobin transcripts using in vitro CRISPR-Cas9 treatment was designed, thus improving parasite transcriptome coverage in low parasitaemia samples. To prove the functionality of the pipeline for both laboratory and field strains, the highest \u00a02-hour resolution RNA-seq transcriptome for\n                      <jats:italic>P. falciparum<\/jats:italic>\n                      3D7 intraerythrocytic life cycle available to\u00a0\u00a0date was generated, and the entire protocol was applied to create the largest transcriptome data from Southeast Asian field isolates.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      Overall, the presented methodology is an inclusive pipeline for generation of good quality transcriptomic data from a diverse range of\n                      <jats:italic>Plasmodium<\/jats:italic>\n                      -infected blood samples with varying parasitaemia and RNA inputs. The flexibility of this pipeline to be adapted to robotic handling will facilitate both small and large-scale future transcriptomic studies in the field of malaria.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12936-020-03436-w","type":"journal-article","created":{"date-parts":[[2020,10,9]],"date-time":"2020-10-09T09:03:00Z","timestamp":1602234180000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":45,"title":["A comprehensive RNA handling and transcriptomics guide for high-throughput processing of Plasmodium blood-stage samples"],"prefix":"10.1186","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1505-5608","authenticated-orcid":false,"given":"Michal","family":"Kucharski","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3461-4767","authenticated-orcid":false,"given":"Jaishree","family":"Tripathi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sourav","family":"Nayak","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lei","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grennady","family":"Wirjanata","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rob W.","family":"van der Pluijm","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mehul","family":"Dhorda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arjen","family":"Dondorp","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9830-8446","authenticated-orcid":false,"given":"Zbynek","family":"Bozdech","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,10,9]]},"reference":[{"key":"3436_CR1","volume-title":"World malaria report 2019","author":"WHO","year":"2020","unstructured":"WHO. World malaria report 2019. Geneva: World Health Organization; 2020."},{"key":"3436_CR2","volume-title":"Guidelines for the treatment of malaria","author":"WHO","year":"2019","unstructured":"WHO. Guidelines for the treatment of malaria. 3rd ed. Geneva: World Health Organization; 2019.","edition":"3"},{"key":"3436_CR3","doi-asserted-by":"publisher","first-page":"411","DOI":"10.1056\/NEJMoa1314981","volume":"371","author":"EA Ashley","year":"2014","unstructured":"Ashley EA, Dhorda M, Fairhurst RM, Amaratunga C, Lim P, Suon S, et al. Spread of artemisinin resistance in Plasmodium falciparum malaria. N Engl J Med. 2014;371:411\u201323.","journal-title":"N Engl J Med"},{"key":"3436_CR4","doi-asserted-by":"publisher","first-page":"1771","DOI":"10.3389\/fimmu.2019.01771","volume":"10","author":"C Penha-Gon\u00e7alves","year":"2019","unstructured":"Penha-Gon\u00e7alves C. Genetics of malaria inflammatory responses: a pathogenesis perspective. Front Immunol. 2019;10:1771.","journal-title":"Front Immunol"},{"key":"3436_CR5","doi-asserted-by":"publisher","first-page":"211ps18","DOI":"10.1126\/scitranslmed.3007432","volume":"5","author":"AJ Cunnington","year":"2013","unstructured":"Cunnington AJ, Walther M, Riley EM. Piecing together the puzzle of severe malaria. Sci Transl Med. 2013;5:211ps18.","journal-title":"Sci Transl Med"},{"key":"3436_CR6","doi-asserted-by":"publisher","first-page":"502","DOI":"10.1038\/nature12216","volume":"498","author":"L Turner","year":"2013","unstructured":"Turner L, Lavstsen T, Berger SS, Wang CW, Petersen JEV, Avril M, et al. Severe malaria is associated with parasite binding to endothelial protein C receptor. Nature. 2013;498:502\u20135.","journal-title":"Nature"},{"key":"3436_CR7","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1046\/j.1365-2958.2003.03570.x","volume":"49","author":"A Salanti","year":"2003","unstructured":"Salanti A, Staalsoe T, Lavstsen T, Jensen ATR, Sowa MPK, Arnot DE, et al. Selective upregulation of a single distinctly structured var gene in chondroitin sulphate A-adhering Plasmodium falciparum involved in pregnancy-associated malaria. Mol Microbiol. 2003;49:179\u201391.","journal-title":"Mol Microbiol"},{"key":"3436_CR8","doi-asserted-by":"publisher","first-page":"e5","DOI":"10.1371\/journal.pbio.0000005","volume":"1","author":"Z Bozdech","year":"2003","unstructured":"Bozdech Z, Llin\u00e1s M, Pulliam BL, Wong ED, Zhu J, DeRisi JL. The transcriptome of the intraerythrocytic developmental cycle of Plasmodium falciparum. PLoS Biol. 2003;1:e5.","journal-title":"PLoS Biol"},{"key":"3436_CR9","doi-asserted-by":"publisher","first-page":"253","DOI":"10.1038\/nature12970","volume":"507","author":"A Sinha","year":"2014","unstructured":"Sinha A, Hughes KR, Modrzynska KK, Otto TD, Pfander C, Dickens NJ, et al. A cascade of DNA-binding proteins for sexual commitment and development in Plasmodium. Nature. 2014;507:253\u20137.","journal-title":"Nature"},{"key":"3436_CR10","doi-asserted-by":"publisher","first-page":"431","DOI":"10.1126\/science.1260403","volume":"347","author":"S Mok","year":"2015","unstructured":"Mok S, Ashley EA, Ferreira PE, Zhu L, Lin Z, Yeo T, et al. Population transcriptomics of human malaria parasites reveals the mechanism of artemisinin resistance. Science. 2015;347:431\u20135.","journal-title":"Science"},{"key":"3436_CR11","doi-asserted-by":"publisher","first-page":"5158","DOI":"10.1038\/s41467-018-07588-x","volume":"9","author":"L Zhu","year":"2018","unstructured":"Zhu L, Tripathi J, Rocamora FM, Miotto O, van der Pluijm R, Voss TS, et al. The origins of malaria artemisinin resistance defined by a genetic and transcriptomic background. Nat Commun. 2018;9:5158.","journal-title":"Nat Commun"},{"key":"3436_CR12","doi-asserted-by":"publisher","first-page":"eaar3619","DOI":"10.1126\/scitranslmed.aar3619","volume":"10","author":"HJ Lee","year":"2018","unstructured":"Lee HJ, Georgiadou A, Walther M, Nwakanma D, Stewart LB, Levin M, et al. Integrated pathogen load and dual transcriptome analysis of systemic host-pathogen interactions in severe malaria. Sci Transl Med. 2018;10:eaar3619.","journal-title":"Sci Transl Med"},{"key":"3436_CR13","doi-asserted-by":"publisher","first-page":"1603","DOI":"10.1086\/522012","volume":"196","author":"A Siau","year":"2007","unstructured":"Siau A, Tour\u00e9 FS, Ouwe-Missi-Oukem-Boyer O, Cic\u00e9ron L, Mahmoudi N, Vaquero C, et al. Whole transcriptome analysis of Plasmodium falciparum field isolates: identification of new pathogenicity factors. J Infect Dis. 2007;196:1603\u201312.","journal-title":"J Infect Dis"},{"key":"3436_CR14","doi-asserted-by":"publisher","first-page":"418","DOI":"10.2307\/3280287","volume":"65","author":"C Lambros","year":"1979","unstructured":"Lambros C, Vanderberg JP. Synchronization of Plasmodium falciparum erythrocytic stages in culture. J Parasitol. 1979;65:418\u201320.","journal-title":"J Parasitol"},{"key":"3436_CR15","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1007\/978-1-62703-026-7_13","volume":"923","author":"Z Bozdech","year":"2012","unstructured":"Bozdech Z, Mok S, Gupta AP. DNA microarray-based genome-wide analyses of Plasmodium parasites. Methods Mol Biol. 2012;923:189\u201321111.","journal-title":"Methods Mol Biol"},{"key":"3436_CR16","doi-asserted-by":"publisher","first-page":"e47","DOI":"10.1093\/nar\/gkv007","volume":"43","author":"G Smyth","year":"2015","unstructured":"Smyth G, Hu Y, Ritchie M, Silver J, Wettenhall J, McCarthy D, et al. limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 2015;43:e47.","journal-title":"Nucleic Acids Res"},{"key":"3436_CR17","doi-asserted-by":"publisher","first-page":"1345","DOI":"10.1016\/S0140-6736(20)30552-3","volume":"395","author":"RW van der Pluijm","year":"2020","unstructured":"van der Pluijm RW, Tripura R, Hoglund RM, Pyae Phyo A, Lek D, Ul Islam A, et al. Triple artemisinin-based combination therapies versus artemisinin-based combination therapies for uncomplicated Plasmodium falciparum malaria: a multicentre, open-label, randomised clinical trial. Lancet. 2020;395:1345\u201360.","journal-title":"Lancet"},{"key":"3436_CR18","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1186\/1471-2199-7-3","volume":"7","author":"A Schroeder","year":"2006","unstructured":"Schroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, et al. The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol. 2006;7:3.","journal-title":"BMC Mol Biol"},{"key":"3436_CR19","doi-asserted-by":"publisher","first-page":"171","DOI":"10.1038\/nprot.2014.006","volume":"9","author":"S Picelli","year":"2014","unstructured":"Picelli S, Faridani OR, Bj\u00f6rklund \u00c5K, Winberg G, Sagasser S, Sandberg R. Full-length RNA-seq from single cells using Smart-seq2. Nat Protoc. 2014;9:171\u201381.","journal-title":"Nat Protoc"},{"key":"3436_CR20","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1186\/s13059-016-0904-5","volume":"17","author":"W Gu","year":"2016","unstructured":"Gu W, Crawford ED, O\u2019Donovan BD, Wilson MR, Chow ED, Retallack H, et al. Depletion of abundant sequences by hybridization (DASH): Using Cas9 to remove unwanted high-abundance species in sequencing libraries and molecular counting applications. Genome Biol. 2016;17:41.","journal-title":"Genome Biol"},{"key":"3436_CR21","doi-asserted-by":"publisher","first-page":"824","DOI":"10.4269\/ajtmh.12-0675","volume":"89","author":"SC Murphy","year":"2013","unstructured":"Murphy SC, Shott JP, Parikh S, Etter P, Prescott WR, Stewart VA. Malaria diagnostics in clinical trials. Am J Trop Med Hyg. 2013;89:824\u201339.","journal-title":"Am J Trop Med Hyg"},{"key":"3436_CR22","doi-asserted-by":"publisher","first-page":"292","DOI":"10.1038\/ng1203-292","volume":"35","author":"H Auer","year":"2003","unstructured":"Auer H, Lyianarachchi S, Newsom D, Klisovic MI, Marcucci U, Kornacker K. Chipping away at the chip bias: RNA degradation in microarray analysis. Nat Genet. 2003;35:292\u20133.","journal-title":"Nat Genet"},{"key":"3436_CR23","doi-asserted-by":"publisher","first-page":"943","DOI":"10.1016\/S1473-3099(19)30392-5","volume":"19","author":"WL Hamilton","year":"2019","unstructured":"Hamilton WL, Amato R, van der Pluijm RW, Jacob CG, Quang HH, Thuy-Nhien NT, et al. Evolution and expansion of multidrug-resistant malaria in southeast Asia: a genomic epidemiology study. Lancet Infect Dis. 2019;19:943\u201351.","journal-title":"Lancet Infect Dis"},{"key":"3436_CR24","doi-asserted-by":"publisher","first-page":"2569","DOI":"10.1007\/s00216-018-0931-z","volume":"410","author":"M Sidstedt","year":"2018","unstructured":"Sidstedt M, Hedman J, Romsos EL, Waitara L, Wads\u00f6 L, Steffen CR, et al. Inhibition mechanisms of hemoglobin, immunoglobulin G, and whole blood in digital and real-time PCR. Anal Bioanal Chem. 2018;410:2569\u201383.","journal-title":"Anal Bioanal Chem"},{"key":"3436_CR25","doi-asserted-by":"publisher","first-page":"581","DOI":"10.1038\/nprot.2006.83","volume":"1","author":"P Chomczynski","year":"2006","unstructured":"Chomczynski P, Sacchi N. The single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction: Twenty-something years on. Nat Protoc. 2006;1:581\u20135.","journal-title":"Nat Protoc"},{"key":"3436_CR26","doi-asserted-by":"publisher","first-page":"1433","DOI":"10.1101\/gr.158980.113","volume":"24","author":"J Yamagishi","year":"2014","unstructured":"Yamagishi J, Natori A, Tolba MEM, Mongan AE, Sugimoto C, Katayama T, et al. Interactive transcriptome analysis of malaria patients and infecting Plasmodium falciparum. Genome Res. 2014;24:1433\u201344.","journal-title":"Genome Res"},{"key":"3436_CR27","doi-asserted-by":"publisher","first-page":"e00071","DOI":"10.1128\/MMBR.00071-17","volume":"82","author":"HJ Lee","year":"2018","unstructured":"Lee HJ, Georgiadou A, Otto TD, Levin M, Coin LJ, Conway DJ, et al. Transcriptomic studies of malaria: a paradigm for investigation of systemic host-pathogen interactions. Microbiol Mol Biol Rev. 2018;82:e00071\u2013e117.","journal-title":"Microbiol Mol Biol Rev"},{"key":"3436_CR28","doi-asserted-by":"publisher","first-page":"3303","DOI":"10.1128\/JCM.01057-14","volume":"52","author":"M Imwong","year":"2014","unstructured":"Imwong M, Hanchana S, Malleret B, R\u00e9nia L, Day NPJ, Dondorp A, et al. High-throughput ultrasensitive molecular techniques for quantifying low-density malaria parasitemias. J Clin Microbiol. 2014;52:3303\u20139.","journal-title":"J Clin Microbiol"},{"key":"3436_CR29","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1186\/s12936-019-2659-4","volume":"18","author":"A Ramaprasad","year":"2019","unstructured":"Ramaprasad A, Subudhi AK, Culleton R, Pain A. A fast and cost-effective microsampling protocol incorporating reduced animal usage for time-series transcriptomics in rodent malaria parasites. Malar J. 2019;18:26.","journal-title":"Malar J"},{"key":"3436_CR30","doi-asserted-by":"publisher","first-page":"442","DOI":"10.1016\/j.ebiom.2019.09.001","volume":"48","author":"R Hoo","year":"2019","unstructured":"Hoo R, Bruske E, Dimonte S, Zhu L, Mordm\u00fcller B, Sim BKL, et al. Transcriptome profiling reveals functional variation in Plasmodium falciparum parasites from controlled human malaria infection studies. EBioMedicine. 2019;48:442\u201352.","journal-title":"EBioMedicine"},{"key":"3436_CR31","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1186\/1475-2875-11-41","volume":"11","author":"M Venkatesan","year":"2012","unstructured":"Venkatesan M, Amaratunga C, Campino S, Auburn S, Koch O, Lim P, et al. Using CF11 cellulose columns to inexpensively and effectively remove human DNA from Plasmodium falciparum-infected whole blood samples. Malar J. 2012;11:41.","journal-title":"Malar J"},{"key":"3436_CR32","doi-asserted-by":"publisher","first-page":"2619","DOI":"10.1126\/science.aaw2619","volume":"365","author":"VM Howick","year":"2019","unstructured":"Howick VM, Russell AJC, Andrews T, Heaton H, Reid AJ, Natarajan K, et al. The Malaria Cell Atlas: single parasite transcriptomes across the complete Plasmodium life cycle. Science. 2019;365:2619.","journal-title":"Science"},{"key":"3436_CR33","doi-asserted-by":"publisher","first-page":"1269","DOI":"10.1021\/acsinfecdis.9b00093","volume":"5","author":"AE Sexton","year":"2019","unstructured":"Sexton AE, Doerig C, Creek DJ, Carvalho TG. Post-genomic approaches to understanding malaria parasite biology: linking genes to biological functions. ACS Infect Dis. 2019;5:1269\u201378.","journal-title":"ACS Infect Dis"},{"key":"3436_CR34","doi-asserted-by":"publisher","first-page":"80","DOI":"10.1016\/j.jviromet.2008.11.022","volume":"157","author":"S Perelle","year":"2009","unstructured":"Perelle S, Cavellini L, Burger C, Blaise-Boisseau S, Hennechart-Collette C, Merle G, et al. Use of a robotic RNA purification protocol based on the NucliSens\u00ae easyMAGTM for real-time RT-PCR detection of hepatitis A virus in bottled water. J Virol Methods. 2009;157:80\u20133.","journal-title":"J Virol Methods"},{"key":"3436_CR35","first-page":"e50356","volume":"76","author":"RC Holmberg","year":"2013","unstructured":"Holmberg RC, Gindlesperger A, Stokes T, Brady D, Thakore N, Belgrader P, et al. High-throughput, automated extraction of DNA and RNA from clinical samples using TruTip technology on common liquid handling robots. J Vis Exp. 2013;76:e50356.","journal-title":"J Vis Exp"},{"key":"3436_CR36","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1177\/2211068211435302","volume":"17","author":"F Kong","year":"2012","unstructured":"Kong F, Yuan L, Zheng YF, Chen W. Automatic liquid handling for life science. J Lab Autom. 2012;17:169\u201385.","journal-title":"J Lab Autom"},{"key":"3436_CR37","doi-asserted-by":"publisher","first-page":"298","DOI":"10.1186\/s12864-020-6631-z","volume":"21","author":"E Berglund","year":"2020","unstructured":"Berglund E, Saarenp\u00e4\u00e4 S, Jemt A, Gruselius J, Larsson L, Bergenstr\u00e5hle L, et al. Automation of spatial transcriptomics library preparation to enable rapid and robust insights into spatial organization of tissues. BMC Genomics. 2020;21:298.","journal-title":"BMC Genomics"}],"container-title":["Malaria Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12936-020-03436-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12936-020-03436-w\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12936-020-03436-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,10,10]],"date-time":"2021-10-10T12:44:07Z","timestamp":1633869847000},"score":1,"resource":{"primary":{"URL":"https:\/\/malariajournal.biomedcentral.com\/articles\/10.1186\/s12936-020-03436-w"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,10,9]]},"references-count":37,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2020,12]]}},"alternative-id":["3436"],"URL":"https:\/\/doi.org\/10.1186\/s12936-020-03436-w","relation":{"has-preprint":[{"id-type":"doi","id":"10.21203\/rs.3.rs-37051\/v1","asserted-by":"object"},{"id-type":"doi","id":"10.21203\/rs.3.rs-37051\/v2","asserted-by":"object"}]},"ISSN":["1475-2875"],"issn-type":[{"value":"1475-2875","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,10,9]]},"assertion":[{"value":"23 June 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 October 2020","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 October 2020","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"All clinical samples used for this study were collected with written informed consent from the patients or their legal guardians. All protocols were approved by the appropriate national ethics committees or the Oxford Tropical Research Ethics Committee.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical approval and consent to participate"}},{"value":"The authors declare that they have no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"363"}}