{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T13:43:52Z","timestamp":1787319832835,"version":"build-2736575974"},"reference-count":43,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2020,9,9]],"date-time":"2020-09-09T00:00:00Z","timestamp":1599609600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2020,9,9]],"date-time":"2020-09-09T00:00:00Z","timestamp":1599609600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"Kenya Medical Research Institute Internal Research Grant","award":["L317"],"award-info":[{"award-number":["L317"]}]}],"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>Prompt diagnosis and effective malaria treatment is a key strategy  in malaria control. However, the recommended diagnostic methods, microscopy and rapid diagnostic tests (RDTs), are not supported by robust quality assurance systems in endemic areas. This study compared the performance of routine RDTs and smear microscopy with a simple molecular-based colorimetric loop-mediated isothermal amplification (LAMP) at two different levels of the health care system in a malaria-endemic area of western Kenya.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Patients presenting with clinical symptoms of malaria at Rota Dispensary (level 2) and Siaya County Referral Hospital (level 4) were enrolled into the study after obtaining written informed consent. Capillary blood was collected to test for malaria by RDT and microscopy at the dispensary and county hospital, and for preparation of blood smears and dried blood spots (DBS) for expert microscopy and real-time polymerase chain reaction (RT-PCR). Results of the routine diagnostic tests were compared with those of malachite green loop-mediated isothermal amplification (MG-LAMP) performed at the two facilities.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>A total of 264 participants were enrolled into the study. At the dispensary level, the positivity rate by RDT, expert microscopy, MG-LAMP and RT-PCR was 37%, 30%, 44% and 42%, respectively, and 42%, 43%, 57% and 43% at the county hospital. Using RT-PCR as the reference test, the sensitivity of RDT and MG-LAMP was 78.1% (CI 67.5\u201386.4) and 82.9% (CI 73.0\u201390.3) at Rota dispensary. At Siaya hospital the sensitivity of routine microscopy and MG-LAMP was 83.3% (CI 65.3\u201394.4) and 93.3% (CI 77.9\u201399.2), respectively. Compared to MG-LAMP, there were 14 false positives and 29 false negatives by RDT at Rota dispensary and 3 false positives and 13 false negatives by routine microscopy at Siaya Hospital.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>MG-LAMP is more sensitive than RDTs and microscopy in the detection of malaria parasites at public health facilities and might be a useful quality control tool in resource-limited settings.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12936-020-03397-0","type":"journal-article","created":{"date-parts":[[2020,9,10]],"date-time":"2020-09-10T07:22:47Z","timestamp":1599722567000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Evaluation of the colorimetric malachite green loop-mediated isothermal amplification (MG-LAMP) assay for the detection of malaria species at two different health facilities in a malaria endemic area of western Kenya"],"prefix":"10.1186","volume":"19","author":[{"given":"James","family":"Gachugia","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Winnie","family":"Chebore","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kephas","family":"Otieno","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caroline Wangari","family":"Ngugi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adano","family":"Godana","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0554-1976","authenticated-orcid":false,"given":"Simon","family":"Kariuki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,9,9]]},"reference":[{"key":"3397_CR1","unstructured":"WHO. World malaria report 2018. Geneva, World Health Organization, 2018."},{"key":"3397_CR2","unstructured":"WHO. Global Technical Strategy for Malaria 2016\u20132030. Geneva, World Health Organization; 2015."},{"key":"3397_CR3","unstructured":"WHO. Malaria. Geneva, World Health Organization, 2017."},{"key":"3397_CR4","doi-asserted-by":"publisher","first-page":"e1001590","DOI":"10.1371\/journal.pmed.1001590","volume":"11","author":"GJ Bastiaens","year":"2014","unstructured":"Bastiaens GJ, Bousema T, Leslie T. Scale-up of malaria rapid diagnostic tests and artemisinin-based combination therapy: challenges and perspectives in sub-Saharan Africa. PLoS Med. 2014;11:e1001590.","journal-title":"PLoS Med"},{"key":"3397_CR5","doi-asserted-by":"publisher","first-page":"34416","DOI":"10.3402\/gha.v9.34416","volume":"9","author":"SP Kachur","year":"2016","unstructured":"Kachur SP. Beyond \u201ctest and treat\u201d \u2013 malaria diagnosis for improved pediatric fever management in sub-Saharan Africa by Emily White Johansson. Glob Health Action. 2016;9:34416.","journal-title":"Glob Health Action"},{"key":"3397_CR6","doi-asserted-by":"publisher","first-page":"233","DOI":"10.1186\/s12936-018-2386-2","volume":"17","author":"G Sori","year":"2018","unstructured":"Sori G, Zewdie O, Tadele G, Samuel A. External quality assessment of malaria microscopy diagnosis in selected health facilities in Western Oromia. Ethiopia Malar J. 2018;17:233.","journal-title":"Ethiopia Malar J"},{"key":"3397_CR7","doi-asserted-by":"publisher","first-page":"221","DOI":"10.1186\/s12936-017-1856-2","volume":"16","author":"E Wanja","year":"2017","unstructured":"Wanja E, Achilla RA, Obare P, Adeny R, Moseti C, Otieno V, et al. Evaluation of a laboratory quality assurance pilot programme for malaria diagnostics in low-transmission areas of Kenya, 2013. Malar J. 2017;16:221.","journal-title":"Malar J"},{"key":"3397_CR8","doi-asserted-by":"publisher","first-page":"371","DOI":"10.1186\/s12936-017-2018-2","volume":"16","author":"F Odhiambo","year":"2017","unstructured":"Odhiambo F, Buff AM, Moranga C, Moseti CM, Wesongah JO, Lowther SA, et al. Factors associated with malaria microscopy diagnostic performance following a pilot quality-assurance programme in health facilities in malaria low-transmission areas of Kenya, 2014. Malar J. 2017;16:371.","journal-title":"Malar J"},{"key":"3397_CR9","doi-asserted-by":"crossref","unstructured":"Jemere KA, Melaku MY, Jemeber TH, Abate MA. Performance evaluation of laboratory professionals on malaria microscopy at health facilities in Bahir Dar city administration, Northwest Ethiopia. PLoS One. 2018;13.","DOI":"10.1371\/journal.pone.0203420"},{"key":"3397_CR10","doi-asserted-by":"publisher","first-page":"14718","DOI":"10.1038\/s41598-017-15031-2","volume":"7","author":"KB Beshir","year":"2017","unstructured":"Beshir KB, Sep\u00falveda 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":"3397_CR11","doi-asserted-by":"crossref","unstructured":"WHO. Malaria rapid diagnostic test performance. Results of WHO product testing of malaria RDTs: Round 3 (2010\u20132011). Geneva, World Health Organization; 2011.","DOI":"10.2471\/TDR.09.978-924-1598071"},{"key":"3397_CR12","unstructured":"WHO. Global Malaria Programme: Policy brief on malaria diagnostics in low-transmission settings. Geneva: World Health Organization; 2014."},{"key":"3397_CR13","doi-asserted-by":"crossref","unstructured":"Berzosa P, de Lucio A, Romay\u2011Barja M, Herrador Z, Gonz\u00e1lez V, Garc\u00eda. L, et al. Comparison of three diagnostic methods (microscopy, RDT, and PCR) for the detection of malaria parasites in representative samples from Equatorial Guinea. Malar J. 2018;17:333.","DOI":"10.1186\/s12936-018-2481-4"},{"key":"3397_CR14","doi-asserted-by":"crossref","unstructured":"Mahmud Y-LL, Meng-Yee L, Mun-Yik F, Jenarun J, Rohela. Loop-mediated isothermal amplification assay for identification of five human Plasmodium species in Malaysia. Am J Trop Med Hyg. 2016;94:336\u20139.","DOI":"10.4269\/ajtmh.15-0569"},{"key":"3397_CR15","unstructured":"Ministry of Health. National guidelines for the diagnosis, treatment and prevention of malaria in Kenya. Nairobi, 2016."},{"key":"3397_CR16","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1186\/s13690-018-0288-y","volume":"76","author":"A Kapesa","year":"2018","unstructured":"Kapesa A, Kweka EJ, Zhou G, Atieli HE, Kamugisha E, Mazigo HD, et al. Utility of passive malaria surveillance in hospitals as a surrogate to community infection transmission dynamics in western Kenya. Arch Public Health. 2018;76:39.","journal-title":"Arch Public Health"},{"key":"3397_CR17","doi-asserted-by":"publisher","first-page":"380","DOI":"10.1186\/1756-3305-7-380","volume":"7","author":"MN Bayoh","year":"2014","unstructured":"Bayoh MN, Walker ED, Kosgei J, Ombok M, Olang GB, Githeko AK, et al. Persistently high estimates of late night, indoor exposure to malaria vectors despite high coverage of insecticide treated nets. Parasit Vectors. 2014;7:380.","journal-title":"Parasit Vectors"},{"key":"3397_CR18","unstructured":"WHO. Basic Malaria Microscopy: Part 1. Learner's Guide. 2nd edition. Geneva, World Health Organization; 2010."},{"key":"3397_CR19","doi-asserted-by":"crossref","unstructured":"Lucchi NW, Ljolje D, Silva-Flannery L, Udhayakumar V. Use of malachite green-loop mediated isothermal amplification for detection of Plasmodium spp. parasites. PLoS One. 2016; 11:e0151437.","DOI":"10.1371\/journal.pone.0151437"},{"key":"3397_CR20","doi-asserted-by":"publisher","first-page":"406","DOI":"10.1186\/s12936-016-1454-8","volume":"15","author":"DB McConnico","year":"2016","unstructured":"McConnico DB, Alessandra EF, Michael CH, John DB, Caitlin C. Development of new malaria diagnostics: matching performance and need. Malar J. 2016;15:406.","journal-title":"Malar J"},{"key":"3397_CR21","doi-asserted-by":"publisher","first-page":"e56677","DOI":"10.1371\/journal.pone.0056677","volume":"8","author":"NW Lucchi","year":"2013","unstructured":"Lucchi NW, Narayanan J, Karell MA, Xayavong M, Kariuki S, DaSilva AJ, et al. Molecular diagnosis of malaria by photo-induced electron transfer fluorogenic primers: PET-PCR. PLoS ONE. 2013;8:e56677.","journal-title":"PLoS ONE"},{"key":"3397_CR22","unstructured":"WHO-FIND-CDC. Methods manual for product testing of malaria rapid diagnostic tests. Geneva, World Health Organization; 2018."},{"key":"3397_CR23","doi-asserted-by":"publisher","first-page":"e001582","DOI":"10.1136\/bmjgh-2019-001582","volume":"4","author":"OJ Watson","year":"2019","unstructured":"Watson OJ, Sumner KM, Janko M, Goel V, Winskill P, Slater HC, et al. False-negative malaria rapid diagnostic test results and their impact on community-based malaria surveys in sub-Saharan Africa. BMJ Glob Health. 2019;4:e001582.","journal-title":"BMJ Glob Health"},{"key":"3397_CR24","doi-asserted-by":"crossref","unstructured":"Shaio P-WL, Dar-Der J, Chia-Tai L, Herodes SR, Virgilio EdR, Lin IF, et al. Application of loop-mediated isothermal amplification for malaria diagnosis during a follow-up study in S\u00e3o Tom\u00e9. Malar J. 2012;11:408.","DOI":"10.1186\/1475-2875-11-408"},{"key":"3397_CR25","doi-asserted-by":"crossref","unstructured":"Bwire GM, Ngasala B, Kilonzi M, Mikomangwa WP, Fatuma F. Felician FF, et al. Diagnostic performance of CareStart\u2122 malaria HRP2\/pLDH test in comparison with standard microscopy for detection of uncomplicated malaria infection among symptomatic patients, Eastern Coast of Tanzania. Malar J. 2019;18:354.","DOI":"10.1186\/s12936-019-2990-9"},{"key":"3397_CR26","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1186\/s12936-019-2711-4","volume":"18","author":"KO Mfuh","year":"2019","unstructured":"Mfuh KO, Achonduh-Atijegbe OA, Bekindaka ON, Esemu LF, Mbakop CD, Gandhi K, et al. A comparison of thick-film microscopy, rapid diagnostic test, and polymerase chain reaction for accurate diagnosis of Plasmodium falciparum malaria. Malar J. 2019;18:73.","journal-title":"Malar J"},{"key":"3397_CR27","doi-asserted-by":"publisher","first-page":"786","DOI":"10.1111\/j.1365-3156.2011.02773.x","volume":"16","author":"JN Waitumbi","year":"2011","unstructured":"Waitumbi JN, Gerlach J, Afonina I, Anyona SB, Koros JN, Siangla J, et al. Malaria prevalence defined by microscopy, antigen detection, DNA amplification and total nucleic acid amplification in a malaria-endemic region during the peak malaria transmission season. Trop Med Int Health. 2011;16:786\u201393.","journal-title":"Trop Med Int Health"},{"key":"3397_CR28","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1186\/s12936-015-0597-3","volume":"14","author":"IM Masanja","year":"2015","unstructured":"Masanja IM, McMorrow ML, Maganga MB, Sumari D, Udhayakumar V, McElroy PD, et al. Quality assurance of malaria rapid diagnostic tests used for routine patient care in rural Tanzania: microscopy versus real-time polymerase chain reaction. Malar J. 2015;14:85.","journal-title":"Malar J"},{"key":"3397_CR29","doi-asserted-by":"publisher","first-page":"456","DOI":"10.1186\/s12936-016-1508-y","volume":"15","author":"EK Wanja","year":"2016","unstructured":"Wanja EK, Moranga N, Hickman C, Johnson M, Moseti J, Anova C, et al. Field evaluation of diagnostic performance of malaria rapid diagnostic tests in western Kenya. Malar J. 2016;15:456.","journal-title":"Malar J"},{"key":"3397_CR30","doi-asserted-by":"publisher","first-page":"e0206848","DOI":"10.1371\/journal.pone.0206848","volume":"13","author":"M Tambo","year":"2018","unstructured":"Tambo M, Mwinga M, Mumbengegwi DR. Loop-mediated isothermal amplification (LAMP) and Polymerase Chain Reaction (PCR) as quality assurance tools for Rapid Diagnostic Test (RDT) malaria diagnosis in Northern Namibia. PLoS ONE. 2018;13:e0206848.","journal-title":"PLoS ONE"},{"key":"3397_CR31","doi-asserted-by":"crossref","unstructured":"ACTwatch Group, Musuva A, Ejersa W, Kiptui R, Memusi D, Abwao E. The malaria testing and treatment landscape in Kenya: results from a nationally representative survey among the public and private sector in 2016. Malar J. 2017;16:494.","DOI":"10.1186\/s12936-017-2089-0"},{"key":"3397_CR32","doi-asserted-by":"publisher","first-page":"764","DOI":"10.1186\/s13104-017-3080-0","volume":"10","author":"HA Hailu","year":"2017","unstructured":"Hailu HA, Shiferaw MB, Demeke L, Derebe MM, Gelaw ZD, Emiru MA, et al. External quality assessment of malaria microscopy diagnosis among public health facilities in West Amhara Region. Ethiopia BMC Res Notes. 2017;10:764.","journal-title":"Ethiopia BMC Res Notes"},{"key":"3397_CR33","doi-asserted-by":"publisher","first-page":"2017","DOI":"10.1093\/infdis\/jit431","volume":"208","author":"KB Beshir","year":"2013","unstructured":"Beshir KB, Sutherland CJ, Sawa P, Drakeley CJ, Okell L, Mweresa CK, et al. Residual Plasmodium falciparum parasitemia in Kenyan children after artemisinin-combination therapy is associated with increased transmission to mosquitoes and parasite recurrence. J Infect Dis. 2013;208:2017\u201324.","journal-title":"J Infect Dis"},{"key":"3397_CR34","unstructured":"WHO. Guidelines for the treatment of malaria. 3rd Edn. Geneva, World Health Organization; 2015."},{"key":"3397_CR35","doi-asserted-by":"crossref","unstructured":"Oriero EC, Okebe J, Jacobs J, Van geertruyden J-P, Nwakanma D, D\u2019Alessandro U. Diagnostic performance of a novel loop-mediated isothermal amplification (LAMP) assay targeting the apicoplast genome for malaria diagnosis in a field setting in sub-Saharan Africa. Malar J. 2015;14:396.","DOI":"10.1186\/s12936-015-0926-6"},{"key":"3397_CR36","doi-asserted-by":"publisher","first-page":"205","DOI":"10.1186\/s12936-015-0731-2","volume":"14","author":"U Morris","year":"2015","unstructured":"Morris U, Khamis M, Aydin-Schmidt B, Abass AK, Msellem MI, Nassor MH, et al. Field deployment of loop-mediated isothermal amplification for centralized mass-screening of asymptomatic malaria in Zanzibar: A pre-elimination setting. Malar J. 2015;14:205.","journal-title":"Malar J"},{"key":"3397_CR37","doi-asserted-by":"publisher","first-page":"98","DOI":"10.1186\/s12936-019-2722-1","volume":"18","author":"HM Kudyba","year":"2019","unstructured":"Kudyba HM, Louzada J, Ljolje D, Kudyba KA, Muralidharan V, Oliveira-Ferreira J, et al. Field evaluation of malaria malachite green loop-mediated isothermal amplification in health posts in Roraima state. Brazil Malar J. 2019;18:98.","journal-title":"Brazil Malar J"},{"key":"3397_CR38","first-page":"11","volume":"5","author":"J Cheaveau","year":"2018","unstructured":"Cheaveau J, Nguyen H, Chow B, Marasinghe D, Mohon AN, Yuan H, et al. Clinical validation of a commercial LAMP test for ruling out malaria in returning travelers: A prospective diagnostic trial. Open Forum Infect Dis. 2018;5:11.","journal-title":"Open Forum Infect Dis"},{"key":"3397_CR39","doi-asserted-by":"publisher","first-page":"373","DOI":"10.1186\/s12936-018-2527-7","volume":"17","author":"JP Vincent","year":"2018","unstructured":"Vincent JP, Komaki-Yasuda K, Iwagami M, Kawai S, Kano S. Combination of PURE-DNA extraction and LAMP-DNA amplification methods for accurate malaria diagnosis on dried blood spots. Malar J. 2018;17:373.","journal-title":"Malar J"},{"key":"3397_CR40","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1186\/s12936-015-0573-y","volume":"14","author":"JC Bj\u00f6rkman","year":"2015","unstructured":"Bj\u00f6rkman JC, Berit A-S, Iveth JG, David B, Elin E, Majda HN, et al. Loop-mediated isothermal amplification (LAMP) for point-of-care detection of asymptomatic low-density malaria parasite carriers in Zanzibar. Malar J. 2015;14:43.","journal-title":"Malar J"},{"key":"3397_CR41","doi-asserted-by":"publisher","first-page":"419","DOI":"10.1186\/s12936-017-2066-7","volume":"16","author":"AN Kabaghe","year":"2017","unstructured":"Kabaghe AN, Phiri MD, Phiri KS, van Vugt M. Challenges in implementing uncomplicated malaria treatment in children: a health facility survey in rural Malawi. Malar J. 2017;16:419.","journal-title":"Malar J"},{"key":"3397_CR42","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1186\/s12941-018-0261-1","volume":"17","author":"MA Diallo","year":"2018","unstructured":"Diallo MA, Diongue K, Seck MC, Ndiaye M, Diallo I, Diedhiou Y, et al. Quality control of malaria microscopy reveals misdiagnosed non-falciparum species and other microscopically detectable pathogens in Senegal. Ann Clin Microbiol Antimicrob. 2018;17:8.","journal-title":"Ann Clin Microbiol Antimicrob"},{"key":"3397_CR43","doi-asserted-by":"publisher","first-page":"446","DOI":"10.1097\/QCO.0000000000000191","volume":"28","author":"PA Zimmerman","year":"2015","unstructured":"Zimmerman PA, Howes RE. Malaria diagnosis for malaria elimination. Curr Opin Infect Dis. 2015;28:446\u201354.","journal-title":"Curr Opin Infect Dis"}],"container-title":["Malaria Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12936-020-03397-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12936-020-03397-0\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12936-020-03397-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,8]],"date-time":"2021-09-08T19:42:56Z","timestamp":1631130176000},"score":1,"resource":{"primary":{"URL":"https:\/\/malariajournal.biomedcentral.com\/articles\/10.1186\/s12936-020-03397-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,9]]},"references-count":43,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2020,12]]}},"alternative-id":["3397"],"URL":"https:\/\/doi.org\/10.1186\/s12936-020-03397-0","relation":{},"ISSN":["1475-2875"],"issn-type":[{"value":"1475-2875","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,9,9]]},"assertion":[{"value":"7 February 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"30 August 2020","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 September 2020","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The protocol was reviewed and approved by the KEMRI Scientific and Ethics Review Unit (SERU) and the Kenyatta University Ethical Review Committee (KU-ERC). All participants or parents\/guardians were informed about the objectives of the study and signed written consent before enrollment into the study.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics 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":"329"}}