{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T03:05:29Z","timestamp":1791947129230,"version":"4.3.4"},"reference-count":38,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,11,14]],"date-time":"2019-11-14T00:00:00Z","timestamp":1573689600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2019,11,14]],"date-time":"2019-11-14T00:00:00Z","timestamp":1573689600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100009139","name":"Deutsches Zentrum f\u00fcr Infektionsforschung","doi-asserted-by":"publisher","award":["TTU01.801"],"award-info":[{"award-number":["TTU01.801"]}],"id":[{"id":"10.13039\/100009139","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009139","name":"Deutsches Zentrum f\u00fcr Infektionsforschung","doi-asserted-by":"publisher","award":["TTU01.801"],"award-info":[{"award-number":["TTU01.801"]}],"id":[{"id":"10.13039\/100009139","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>We describe DAMIAN, an open source bioinformatics tool designed for the identification of pathogenic microorganisms in diagnostic samples. By using authentic clinical samples and comparing our results to those from established analysis pipelines as well as conventional diagnostics, we demonstrate that DAMIAN rapidly identifies pathogens in different diagnostic entities, and accurately classifies viral agents down to the strain level. We furthermore show that DAMIAN is able to assemble full-length viral genomes even in samples co-infected with multiple virus strains, an ability which is of considerable advantage for the investigation of outbreak scenarios. While DAMIAN, similar to other pipelines, analyzes single samples to perform classification of sequences according to their likely taxonomic origin, it also includes a tool for cohort-based analysis. This tool uses cross-sample comparisons to identify sequence signatures that are frequently present in a sample group of interest (e.g., a disease-associated cohort), but occur less frequently in control cohorts. As this approach does not require homology searches in databases, it principally allows the identification of not only known, but also completely novel pathogens. Using samples from a meningitis outbreak, we demonstrate the feasibility of this approach in identifying enterovirus as the causative agent.<\/jats:p>","DOI":"10.1038\/s41598-019-52881-4","type":"journal-article","created":{"date-parts":[[2019,11,14]],"date-time":"2019-11-14T11:02:42Z","timestamp":1573729362000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["DAMIAN: an open source bioinformatics tool for fast, systematic and cohort based analysis of microorganisms in diagnostic samples"],"prefix":"10.1038","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5993-7709","authenticated-orcid":false,"given":"Malik","family":"Alawi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lia","family":"Burkhardt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Indenbirken","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kerstin","family":"Reumann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maximilian","family":"Christopeit","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolaus","family":"Kr\u00f6ger","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc","family":"L\u00fctgehetmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Aepfelbacher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicole","family":"Fischer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0940-7045","authenticated-orcid":false,"given":"Adam","family":"Grundhoff","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,11,14]]},"reference":[{"key":"52881_CR1","doi-asserted-by":"publisher","first-page":"ofy257","DOI":"10.1093\/ofid\/ofy257","volume":"5","author":"T Basein","year":"2018","unstructured":"Basein, T. et al. Microbial Identification Using DNA Target Amplification and Sequencing: Clinical Utility and Impact on Patient Management. Open forum infectious diseases 5, ofy257, https:\/\/doi.org\/10.1093\/ofid\/ofy257 (2018).","journal-title":"Open forum infectious diseases"},{"key":"52881_CR2","doi-asserted-by":"publisher","first-page":"1686","DOI":"10.1128\/JCM.02664-15","volume":"54","author":"LF Westblade","year":"2016","unstructured":"Westblade, L. F. et al. Role of Clinicogenomics in Infectious Disease Diagnostics and Public Health Microbiology. Journal of clinical microbiology 54, 1686\u20131693, https:\/\/doi.org\/10.1128\/JCM.02664-15 (2016).","journal-title":"Journal of clinical microbiology"},{"key":"52881_CR3","doi-asserted-by":"publisher","first-page":"1245","DOI":"10.1093\/cid\/cir692","volume":"53","author":"SK Rampini","year":"2011","unstructured":"Rampini, S. K. et al. Broad-range 16S rRNA gene polymerase chain reaction for diagnosis of culture-negative bacterial infections. Clinical infectious diseases: an official publication of the Infectious Diseases Society of America 53, 1245\u20131251, https:\/\/doi.org\/10.1093\/cid\/cir692 (2011).","journal-title":"Clinical infectious diseases: an official publication of the Infectious Diseases Society of America"},{"key":"52881_CR4","doi-asserted-by":"publisher","first-page":"e65226","DOI":"10.1371\/journal.pone.0065226","volume":"8","author":"SJ Salipante","year":"2013","unstructured":"Salipante, S. J. et al. Rapid 16S rRNA next-generation sequencing of polymicrobial clinical samples for diagnosis of complex bacterial infections. PloS one 8, e65226, https:\/\/doi.org\/10.1371\/journal.pone.0065226 (2013).","journal-title":"PloS one"},{"key":"52881_CR5","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-25129-w","volume":"8","author":"K Wagner","year":"2018","unstructured":"Wagner, K., Springer, B., Pires, V. P. & Keller, P. M. Molecular detection of fungal pathogens in clinical specimens by 18S rDNA high-throughput screening in comparison to ITS PCR and culture. Scientific reports 8, 6964, https:\/\/doi.org\/10.1038\/s41598-018-25129-w (2018).","journal-title":"Scientific reports"},{"key":"52881_CR6","doi-asserted-by":"publisher","first-page":"1072","DOI":"10.3201\/eid2006.131526","volume":"20","author":"N Fischer","year":"2014","unstructured":"Fischer, N. et al. Rapid metagenomic diagnostics for suspected outbreak of severe pneumonia. Emerging infectious diseases 20, 1072\u20131075, https:\/\/doi.org\/10.3201\/eid2006.131526 (2014).","journal-title":"Emerging infectious diseases"},{"key":"52881_CR7","doi-asserted-by":"publisher","first-page":"1502","DOI":"10.1001\/jama.2013.3231","volume":"309","author":"NJ Loman","year":"2013","unstructured":"Loman, N. J. et al. A culture-independent sequence-based metagenomics approach to the investigation of an outbreak of Shiga-toxigenic Escherichia coli O104:H4. Jama 309, 1502\u20131510, https:\/\/doi.org\/10.1001\/jama.2013.3231 (2013).","journal-title":"Jama"},{"key":"52881_CR8","doi-asserted-by":"publisher","first-page":"919","DOI":"10.1093\/cid\/ciu912","volume":"60","author":"SN Naccache","year":"2015","unstructured":"Naccache, S. N. et al. Diagnosis of neuroinvasive astrovirus infection in an immunocompromised adult with encephalitis by unbiased next-generation sequencing. Clinical infectious diseases: an official publication of the Infectious Diseases Society of America 60, 919\u2013923, https:\/\/doi.org\/10.1093\/cid\/ciu912 (2015).","journal-title":"Clinical infectious diseases: an official publication of the Infectious Diseases Society of America"},{"key":"52881_CR9","doi-asserted-by":"publisher","first-page":"2408","DOI":"10.1056\/NEJMoa1401268","volume":"370","author":"MR Wilson","year":"2014","unstructured":"Wilson, M. R. et al. Actionable diagnosis of neuroleptospirosis by next-generation sequencing. The New England journal of medicine 370, 2408\u20132417, https:\/\/doi.org\/10.1056\/NEJMoa1401268 (2014).","journal-title":"The New England journal of medicine"},{"key":"52881_CR10","doi-asserted-by":"publisher","first-page":"341","DOI":"10.1038\/s41576-019-0113-7","volume":"20","author":"CY Chiu","year":"2019","unstructured":"Chiu, C. Y. & Miller, S. A. Clinical metagenomics. Nat Rev Genet 20, 341\u2013355, https:\/\/doi.org\/10.1038\/s41576-019-0113-7 (2019).","journal-title":"Nat Rev Genet"},{"key":"52881_CR11","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-016-0969-1","volume":"17","author":"S Flygare","year":"2016","unstructured":"Flygare, S. et al. Taxonomer: an interactive metagenomics analysis portal for universal pathogen detection and host mRNA expression profiling. Genome biology 17, 111, https:\/\/doi.org\/10.1186\/s13059-016-0969-1 (2016).","journal-title":"Genome biology"},{"key":"52881_CR12","doi-asserted-by":"publisher","first-page":"831","DOI":"10.1101\/gr.238170.118","volume":"29","author":"S Miller","year":"2019","unstructured":"Miller, S. et al. Laboratory validation of a clinical metagenomic sequencing assay for pathogen detection in cerebrospinal fluid. Genome research 29, 831\u2013842, https:\/\/doi.org\/10.1101\/gr.238170.118 (2019).","journal-title":"Genome research"},{"key":"52881_CR13","doi-asserted-by":"publisher","first-page":"776","DOI":"10.5858\/arpa.2016-0539-RA","volume":"141","author":"R Schlaberg","year":"2017","unstructured":"Schlaberg, R. et al. Validation of Metagenomic Next-Generation Sequencing Tests for Universal Pathogen Detection. Archives of pathology &. laboratory medicine 141, 776\u2013786, https:\/\/doi.org\/10.5858\/arpa.2016-0539-RA (2017).","journal-title":"laboratory medicine"},{"key":"52881_CR14","doi-asserted-by":"publisher","first-page":"E976","DOI":"10.1073\/pnas.1317064111","volume":"111","author":"SN Naccache","year":"2014","unstructured":"Naccache, S. N., Hackett, J. Jr., Delwart, E. L. & Chiu, C. Y. Concerns over the origin of NIH-CQV, a novel virus discovered in Chinese patients with seronegative hepatitis. Proceedings of the National Academy of Sciences of the United States of America 111, E976, https:\/\/doi.org\/10.1073\/pnas.1317064111 (2014).","journal-title":"Proceedings of the National Academy of Sciences of the United States of America"},{"key":"52881_CR15","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2014-15-3-r46","volume":"15","author":"DE Wood","year":"2014","unstructured":"Wood, D. E. & Salzberg, S. L. Kraken: ultrafast metagenomic sequence classification using exact alignments. Genome biology 15, R46, https:\/\/doi.org\/10.1186\/gb-2014-15-3-r46 (2014).","journal-title":"Genome biology"},{"key":"52881_CR16","doi-asserted-by":"publisher","first-page":"1721","DOI":"10.1101\/gr.150151.112","volume":"23","author":"OE Francis","year":"2013","unstructured":"Francis, O. E. et al. Pathoscope: species identification and strain attribution with unassembled sequencing data. Genome research 23, 1721\u20131729, https:\/\/doi.org\/10.1101\/gr.150151.112 (2013).","journal-title":"Genome research"},{"key":"52881_CR17","doi-asserted-by":"publisher","first-page":"2930","DOI":"10.1093\/bioinformatics\/btv317","volume":"31","author":"S Morfopoulou","year":"2015","unstructured":"Morfopoulou, S. & Plagnol, V. Bayesian mixture analysis for metagenomic community profiling. Bioinformatics 31, 2930\u20132938, https:\/\/doi.org\/10.1093\/bioinformatics\/btv317 (2015).","journal-title":"Bioinformatics"},{"key":"52881_CR18","doi-asserted-by":"publisher","first-page":"W275","DOI":"10.1093\/nar\/gkm365","volume":"35","author":"G Lu","year":"2007","unstructured":"Lu, G., Rowley, T., Garten, R. & Donis, R. O. FluGenome: a web tool for genotyping influenza A virus. Nucleic acids research 35, W275\u2013279, https:\/\/doi.org\/10.1093\/nar\/gkm365 (2007).","journal-title":"Nucleic acids research"},{"key":"52881_CR19","doi-asserted-by":"publisher","first-page":"612","DOI":"10.1016\/j.ijmm.2015.08.010","volume":"305","author":"M Hohne","year":"2015","unstructured":"Hohne, M., Niendorf, S., Mas Marques, A. & Bock, C. T. Use of sequence analysis of the P2 domain for characterization of norovirus strains causing a large multistate outbreak of norovirus gastroenteritis in Germany 2012. Int J Med Microbiol 305, 612\u2013618, https:\/\/doi.org\/10.1016\/j.ijmm.2015.08.010 (2015).","journal-title":"Int J Med Microbiol"},{"key":"52881_CR20","doi-asserted-by":"publisher","DOI":"10.1007\/s12560-013-9118-0","author":"D Made","year":"2013","unstructured":"Made, D., Trubner, K., Neubert, E., Hohne, M. & Johne, R. Detection and Typing of Norovirus from Frozen Strawberries Involved in a Large-Scale Gastroenteritis Outbreak in Germany. Food and environmental virology. https:\/\/doi.org\/10.1007\/s12560-013-9118-0 (2013).","journal-title":"Food and environmental virology"},{"key":"52881_CR21","doi-asserted-by":"publisher","first-page":"246203","DOI":"10.1155\/2015\/246203","volume":"2015","author":"C Vincent","year":"2015","unstructured":"Vincent, C., Mehrotra, S., Loo, V. G., Dewar, K. & Manges, A. R. Excretion of Host DNA in Feces Is Associated with Risk of Clostridium difficile Infection. J Immunol Res 2015, 246203, https:\/\/doi.org\/10.1155\/2015\/246203 (2015).","journal-title":"J Immunol Res"},{"issue":"2","key":"52881_CR22","doi-asserted-by":"publisher","first-page":"53","DOI":"10.3390\/v8020053","volume":"8","author":"Jens Friis-Nielsen","year":"2016","unstructured":"Friis-Nielsen, J. et al. Identification of Known and Novel Recurrent Viral Sequences in Data from Multiple Patients and Multiple Cancers. Viruses 8, https:\/\/doi.org\/10.3390\/v8020053 (2016).","journal-title":"Viruses"},{"key":"52881_CR23","doi-asserted-by":"publisher","first-page":"1398","DOI":"10.1128\/JVI.03206-13","volume":"88","author":"H Smuts","year":"2014","unstructured":"Smuts, H., Kew, M., Khan, A. & Korsman, S. Novel hybrid parvovirus-like virus, NIH-CQV\/PHV, contaminants in silica column-based nucleic acid extraction kits. Journal of virology 88, 1398, https:\/\/doi.org\/10.1128\/JVI.03206-13 (2014).","journal-title":"Journal of virology"},{"key":"52881_CR24","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-03997-y","volume":"7","author":"T Gunther","year":"2017","unstructured":"Gunther, T. et al. Recovery of the first full-length genome sequence of a parapoxvirus directly from a clinical sample. Scientific reports 7, 3734, https:\/\/doi.org\/10.1038\/s41598-017-03997-y (2017).","journal-title":"Scientific reports"},{"key":"52881_CR25","doi-asserted-by":"publisher","first-page":"2114","DOI":"10.1093\/bioinformatics\/btu170","volume":"30","author":"AM Bolger","year":"2014","unstructured":"Bolger, A. M., Lohse, M. & Usadel, B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30, 2114\u20132120, https:\/\/doi.org\/10.1093\/bioinformatics\/btu170 (2014).","journal-title":"Bioinformatics"},{"key":"52881_CR26","doi-asserted-by":"publisher","DOI":"10.1186\/s13040-014-0034-0","volume":"8","author":"WB Langdon","year":"2015","unstructured":"Langdon, W. B. Performance of genetic programming optimised Bowtie2 on genome comparison and analytic testing (GCAT) benchmarks. BioData Min 8, 1, https:\/\/doi.org\/10.1186\/s13040-014-0034-0 (2015).","journal-title":"BioData Min"},{"key":"52881_CR27","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2009-10-3-r25","volume":"10","author":"B Langmead","year":"2009","unstructured":"Langmead, B., Trapnell, C., Pop, M. & Salzberg, S. L. Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome biology 10, R25, https:\/\/doi.org\/10.1186\/gb-2009-10-3-r25 (2009).","journal-title":"Genome biology"},{"key":"52881_CR28","doi-asserted-by":"publisher","first-page":"1420","DOI":"10.1093\/bioinformatics\/bts174","volume":"28","author":"Y Peng","year":"2012","unstructured":"Peng, Y., Leung, H. C., Yiu, S. M. & Chin, F. Y. IDBA-UD: a de novo assembler for single-cell and metagenomic sequencing data with highly uneven depth. Bioinformatics 28, 1420\u20131428, https:\/\/doi.org\/10.1093\/bioinformatics\/bts174 (2012).","journal-title":"Bioinformatics"},{"key":"52881_CR29","doi-asserted-by":"publisher","first-page":"e1002195","DOI":"10.1371\/journal.pcbi.1002195","volume":"7","author":"SR Eddy","year":"2011","unstructured":"Eddy, S. R. Accelerated Profile HMM Searches. PLoS computational biology 7, e1002195, https:\/\/doi.org\/10.1371\/journal.pcbi.1002195 (2011).","journal-title":"PLoS computational biology"},{"key":"52881_CR30","doi-asserted-by":"publisher","first-page":"D427","DOI":"10.1093\/nar\/gky995","volume":"47","author":"S El-Gebali","year":"2019","unstructured":"El-Gebali, S. et al. The Pfam protein families database in 2019. Nucleic acids research 47, D427\u2013D432, https:\/\/doi.org\/10.1093\/nar\/gky995 (2019).","journal-title":"Nucleic acids research"},{"key":"52881_CR31","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-10-421","volume":"10","author":"C Camacho","year":"2009","unstructured":"Camacho, C. et al. BLAST+: architecture and applications. BMC bioinformatics 10, 421, https:\/\/doi.org\/10.1186\/1471-2105-10-421 (2009).","journal-title":"BMC bioinformatics"},{"key":"52881_CR32","doi-asserted-by":"publisher","first-page":"58","DOI":"10.1016\/j.jcv.2007.11.016","volume":"42","author":"U Dierssen","year":"2008","unstructured":"Dierssen, U., Rehren, F., Henke-Gendo, C., Harste, G. & Heim, A. Rapid routine detection of enterovirus RNA in cerebrospinal fluid by a one-step real-time RT-PCR assay. Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 42, 58\u201364, https:\/\/doi.org\/10.1016\/j.jcv.2007.11.016 (2008).","journal-title":"Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology"},{"key":"52881_CR33","doi-asserted-by":"publisher","first-page":"2238","DOI":"10.1128\/JCM.02495-14","volume":"53","author":"N Fischer","year":"2015","unstructured":"Fischer, N. et al. Evaluation of Unbiased Next-Generation Sequencing of RNA (RNA-seq) as a Diagnostic Method in Influenza Virus-Positive Respiratory Samples. Journal of clinical microbiology 53, 2238\u20132250, https:\/\/doi.org\/10.1128\/JCM.02495-14 (2015).","journal-title":"Journal of clinical microbiology"},{"key":"52881_CR34","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1016\/j.jcv.2011.04.010","volume":"51","author":"RR Jansen","year":"2011","unstructured":"Jansen, R. R. et al. Development and evaluation of a four-tube real time multiplex PCR assay covering fourteen respiratory viruses, and comparison to its corresponding single target counterparts. Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 51, 179\u2013185, https:\/\/doi.org\/10.1016\/j.jcv.2011.04.010 (2011).","journal-title":"Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology"},{"key":"52881_CR35","doi-asserted-by":"publisher","first-page":"1674","DOI":"10.1128\/JVI.02109-09","volume":"84","author":"L Li","year":"2010","unstructured":"Li, L. et al. Multiple diverse circoviruses infect farm animals and are commonly found in human and chimpanzee feces. Journal of virology 84, 1674\u20131682, https:\/\/doi.org\/10.1128\/JVI.02109-09 (2010).","journal-title":"Journal of virology"},{"key":"52881_CR36","unstructured":"Panning, M. et al. Detection of influenza A(H1N1)v virus by real-time RT-PCR. Euro surveillance: bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin 14 (2009)."},{"key":"52881_CR37","doi-asserted-by":"publisher","first-page":"2868","DOI":"10.1128\/JCM.06752-11","volume":"50","author":"M Schibler","year":"2012","unstructured":"Schibler, M. et al. Critical analysis of rhinovirus RNA load quantification by real-time reverse transcription-PCR. Journal of clinical microbiology 50, 2868\u20132872, https:\/\/doi.org\/10.1128\/JCM.06752-11 (2012).","journal-title":"Journal of clinical microbiology"},{"key":"52881_CR38","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1016\/S1386-6532(03)00122-7","volume":"29","author":"CL Ward","year":"2004","unstructured":"Ward, C. L. et al. Design and performance testing of quantitative real time PCR assays for influenza A and B viral load measurement. Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 29, 179\u2013188, https:\/\/doi.org\/10.1016\/S1386-6532(03)00122-7 (2004).","journal-title":"Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-52881-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-52881-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-52881-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,17]],"date-time":"2022-12-17T16:01:10Z","timestamp":1671292870000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-52881-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,11,14]]},"references-count":38,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["52881"],"URL":"https:\/\/doi.org\/10.1038\/s41598-019-52881-4","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,11,14]]},"assertion":[{"value":"10 May 2019","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"24 October 2019","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"14 November 2019","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"16841"}}