{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,12]],"date-time":"2026-07-12T05:07:41Z","timestamp":1783832861516,"version":"3.55.0"},"reference-count":39,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T00:00:00Z","timestamp":1753833600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T00:00:00Z","timestamp":1753833600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2025,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>This study introduces a new procedure for antiviral resistance analysis and genetic classification of human cytomegalovirus (HCMV) using next\u2010generation sequencing (NGS) adapted to existing methodologies, aiming for more targets due to the recent use of new antivirals. It expands the classical investigation of mutations in UL54 and UL97 genes, associated with resistance to (val)ganciclovir, foscarnet and cidofovir, to include UL27, UL56 and UL89 genes, which target newer antivirals like maribavir and letermovir. Additionally, it includes the genetic analysis of UL55 (glycoprotein B) for genotype classification. This new methodology involves multiplex\u2010PCR for DNA enrichment, followed by NGS using Illumina MiSeq platform and data analysis through an in\u2010house pipeline. Several validations were performed by firstly using genome sequence from wild\u2010type sensitive reference strain (AD\u2010169), secondly comparing to previously characterized samples by Sanger, and lastly the use of external quality controls. A new NGS technique based on amplicons approach has been developed. Validation using wild\u2010type control material showed 100% identity with the reference genome across all replicates. Only one minority variant was detected in one replicate. Compared to Sanger sequencing, NGS revealed additional low\u2010frequency mutations not detected by Sanger, without impacting resistance interpretation. The method also performed reliably in external quality assessment controls. Moreover, the detection limit of the technique was established at 17\u2009894.60\u2009IU\/mL. Finally, this approach enabled the identification of HCMV genotypes. This approach improves the monitoring of antiviral resistance and viral diversity, enhancing early clinical decision\u2010making in immunocompromised patients.<\/jats:p>","DOI":"10.1002\/jmv.70523","type":"journal-article","created":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T11:05:15Z","timestamp":1753873515000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A New Next\u2010Generation Sequencing Approach in Human Cytomegalovirus for the Identification of Antiviral Resistance Mutations and Genotypic Classification"],"prefix":"10.1002","volume":"97","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8015-5170","authenticated-orcid":false,"given":"Maria","family":"Arnedo\u2010Mu\u00f1oz","sequence":"first","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"},{"name":"Department of Genetics and Microbiology Universitat Aut\u00f2noma de Barcelona (UAB) Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ignasi","family":"Prats\u2010M\u00e9ndez","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandra","family":"G\u00f3nzalez\u2010S\u00e1nchez","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Carmen","family":"Mart\u00edn","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ariadna","family":"Rando\u2010Segura","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"},{"name":"CIBER Hepatic and Digestive Diseases (CIBERehd) Instituto Carlos III Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ibai Los","family":"Arcos","sequence":"additional","affiliation":[{"name":"Infectious Diseases Department, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Bellaterra Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en red de Enfermedades Infecciosas CIBERINFEC Instituto Carlos III Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia","family":"Nadal\u2010Bar\u00f3n","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Narc\u00eds","family":"Saubi","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalia","family":"Mendoza\u2010Palomar","sequence":"additional","affiliation":[{"name":"Pediatric Infectious Diseases and Immunodeficiencies Unit Hospital Infantil. Vall d\u2032Hebron Barcelona Hospital Campus, Institut de Recerca Vall d\u2032Hebron Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marta","family":"Bernat\u2010Sole","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adaia","family":"Albasanz\u2010Puig","sequence":"additional","affiliation":[{"name":"Infectious Diseases Department, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Bellaterra Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Pi\u00f1ana","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristina","family":"Andr\u00e9s","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susana","family":"Melendo\u2010P\u00e9rez","sequence":"additional","affiliation":[{"name":"Pediatric Infectious Diseases and Immunodeficiencies Unit Hospital Infantil. Vall d\u2032Hebron Barcelona Hospital Campus, Institut de Recerca Vall d\u2032Hebron Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nieves","family":"Larrosa","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en red de Enfermedades Infecciosas CIBERINFEC Instituto Carlos III Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9s","family":"Ant\u00f3n","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en red de Enfermedades Infecciosas CIBERINFEC Instituto Carlos III Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juliana","family":"Esperalba","sequence":"additional","affiliation":[{"name":"Microbiology Department, Vall d\u2032Hebron Institut de Recerca (VHIR), Vall d\u2032Hebron Hospital Universitari, Vall d\u2032Hebron Barcelona Hospital Campus Universitat Aut\u00f2noma de Barcelona Barcelona Spain"},{"name":"Department of Genetics and Microbiology Universitat Aut\u00f2noma de Barcelona (UAB) Barcelona Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en red de Enfermedades Infecciosas CIBERINFEC Instituto Carlos III Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,7,30]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/rmv.2034"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.eimc.2009.07.008"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1093\/bmb\/ldz031"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2020.104711"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.78.13.7124-7130.2004"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41409-023-01920-w"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00611-13"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1089\/wound.2012.0379"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.24222"},{"key":"e_1_2_9_11_1","doi-asserted-by":"crossref","unstructured":"C.Kent A. D.Smith J.Tyson et al. PrimalScheme: Open\u2010Source Community Resources for Low\u2010Cost Viral Genome Sequencing. Published online December 22 2024 https:\/\/doi.org\/10.1101\/2024.12.20.629611.","DOI":"10.1101\/2024.12.20.629611"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btu170"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1923"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/cpbi.102"},{"key":"e_1_2_9_15_1","unstructured":"S. F.AltschuP W.Gish W.Miller E. W.Myers andD. J.LipmanBasic Local Alignment Search Tool."},{"key":"e_1_2_9_16_1","doi-asserted-by":"crossref","unstructured":"H.LiMinimap2: Pairwise Alignment for Nucleotide Sequences. Birol I. ed. Bioinformatics.2018;34(18):3094\u20133100 https:\/\/doi.org\/10.1093\/bioinformatics\/bty191.","DOI":"10.1093\/bioinformatics\/bty191"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1093\/gigascience\/giab008"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks918"},{"key":"e_1_2_9_19_1","unstructured":"I.Prats\u2010M\u00e9ndezminMutFinder. Published online September 17 2024 https:\/\/github.com\/ValldHebron-Bioinformatics\/minMutFinder."},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2024.106016"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1093\/bib\/bbs017"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/msr121"},{"key":"e_1_2_9_23_1","doi-asserted-by":"crossref","unstructured":"H.Gourl\u00e9 O.Karlsson\u2010Lindsj\u00f6 J.Hayer andE.Bongcam\u2010RudloffSimulating Illumina Metagenomic Data With InSilicoSeq. Hancock J. ed. Bioinformatics.2019;35(3):521\u2013522 https:\/\/doi.org\/10.1093\/bioinformatics\/bty630.","DOI":"10.1093\/bioinformatics\/bty630"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.00829-23"},{"key":"e_1_2_9_25_1","unstructured":"J.Xu \u201cHuman Cytomegalovirus Envelope Glycoprotein B H and N Polymorphisms Among Infants of Shanghai Area in China.\u201d"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01341-08"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/cis1035"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2016-06-688432"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.3389\/ti.2023.11785"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2023.105623"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1034\/j.1600-6143.2001.10403.x"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.3390\/tropicalmed9020049"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.49.3.873-883.2005"},{"key":"e_1_2_9_34_1","doi-asserted-by":"publisher","DOI":"10.1111\/tid.12241"},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0219701"},{"key":"e_1_2_9_36_1","first-page":"321","volume-title":"Methods in Molecular Biology","author":"Nagy P. L.","year":"2018"},{"key":"e_1_2_9_37_1","doi-asserted-by":"publisher","DOI":"10.20892\/j.issn.2095-3941.2018.0055"},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro.2016.182"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.140123497"},{"key":"e_1_2_9_40_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkz196"}],"container-title":["Journal of Medical Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmv.70523","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/jmv.70523","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmv.70523","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,12]],"date-time":"2026-07-12T05:03:07Z","timestamp":1783832587000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jmv.70523"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,30]]},"references-count":39,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2025,8]]}},"alternative-id":["10.1002\/jmv.70523"],"URL":"https:\/\/doi.org\/10.1002\/jmv.70523","archive":["Portico"],"relation":{},"ISSN":["0146-6615","1096-9071"],"issn-type":[{"value":"0146-6615","type":"print"},{"value":"1096-9071","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,30]]},"assertion":[{"value":"2025-02-12","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-07-19","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-07-30","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70523"}}