title: Genotypic resistance of cytomegalovirus to antivirals in hematopoietic stem cell transplant recipients from Portugal: A retrospective study.
abstract: The aim of this study was to characterize Human Cytomegalovirus (HCMV) drug resistance mutations in UL97 and UL54 genes in allogeneic hematopoietic stem cell transplant (allo-HSCT) recipients in Portugal. We have performed a retrospective study with 22 patients from a cohort of patients with different haematological malignancies submitted to allo-HSCT between 2010 and 2014. Patients were selected according to clinical and laboratory data of HCMV infection and management. HCMV resistance mutations were characterized by sequencing of UL97 and UL54 genes. Sequence data were compared with: 1) HCMV genome reference strain AD169; and also 2) UL97 from Merlin strain (GenBank: AY446894.2), and UL54 from TB40/E strain (GenBank: ABV71585.1). Resistance mutations were identified in seven patients (32%): five harboured resistance mutations in UL97: A594V (n = 2), C592G (n = 1), L595W (n = 1), and C603W (n = 1); and two harboured resistance mutations in UL54: P522S and L957F, one in each patient. Several natural polymorphisms and unknown mutations were found in both UL97 and UL54, with the majority of the patients harbouring more than one unknown mutation in UL97 but only one in UL54. No simultaneous mutations were found. This is the first study in Portugal to characterize HCMV UL97 and UL54 sequences and to identify HCMV drug-resistance mutations in allo-HSCT patients. The UL97 resistance mutations found were amongst the most frequent resistant mutations, while UL54 L957F mutation was here reported for the first time in a clinical specimen. This information provides important information regarding HCMV strains and antiviral resistance in our population.
authors.affiliation: Molecular Oncology and Viral Pathology Group, Research Centre (CI-IPOP), Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal; Faculty of Medicine of University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319, Porto, Portugal.
authors.forename: Ana Bela
authors.initials: AB
authors.lastname: Campos
authors.affiliation: Molecular Oncology and Viral Pathology Group, Research Centre (CI-IPOP), Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal; Virology Service, Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal; Faculty of Medicine of University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319, Porto, Portugal.
authors.forename: Joana
authors.initials: J
authors.lastname: Ribeiro
authors.affiliation: Bone Marrow Transplantation Unit, Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal.
authors.forename: Carlos
authors.initials: C
authors.lastname: Pinho Vaz
authors.affiliation: Bone Marrow Transplantation Unit, Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal.
authors.forename: Fernando
authors.initials: F
authors.lastname: Campilho
authors.affiliation: Bone Marrow Transplantation Unit, Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal.
authors.forename: Rosa
authors.initials: R
authors.lastname: Branca
authors.affiliation: Bone Marrow Transplantation Unit, Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal.
authors.forename: António
authors.initials: A
authors.lastname: Campos
authors.affiliation: Virology Service, Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal.
authors.forename: Inês
authors.initials: I
authors.lastname: Baldaque
authors.affiliation: Molecular Oncology and Viral Pathology Group, Research Centre (CI-IPOP), Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal; Virology Service, Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal; Research Department, Portuguese League Against Cancer (LPCC-NRNorte), Estrada Interior da Circunvalação 6657, 4200, Porto, Portugal.
authors.forename: Rui
authors.initials: R
authors.lastname: Medeiros
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, CR7, Centre d'Immunologie et des Maladies Infectieuses (CIMI-Paris), F-75013, Paris, France; INSERM, U1135, CIMI-Paris, F-75013, Paris, France; AP-HP, Hôpitaux Universitaires La Pitié-Salpêtrière - Charles Foix, Service de Virologie, F-75013, Paris, France.
authors.forename: David
authors.initials: D
authors.lastname: Boutolleau
authors.affiliation: Molecular Oncology and Viral Pathology Group, Research Centre (CI-IPOP), Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal; Virology Service, Portuguese Oncology Institute of Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal. Electronic address: hugomls@gmail.com.
authors.forename: Hugo
authors.initials: H
authors.lastname: Sousa
mesh_terms: D000293:Adolescent; D000328:Adult; D000998:Antiviral Agents / Q000494:pharmacology*; D002648:Child; D002675:Child, Preschool; D003587:Cytomegalovirus / Q000187:drug effects* / Q000235:genetics*; D003586:Cytomegalovirus Infections / Q000453:epidemiology / Q000821:virology; D004279:DNA, Viral / Q000187:drug effects; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D024882:Drug Resistance, Viral / Q000235:genetics*; D005260:Female; D015774:Ganciclovir / Q000494:pharmacology; D005838:Genotype; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D009154:Mutation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D011110:Polymorphism, Genetic; D011174:Portugal / Q000453:epidemiology; D012189:Retrospective Studies; D017422:Sequence Analysis, DNA; D066027:Transplant Recipients; D014764:Viral Proteins / Q000235:genetics; D055815:Young Adult
chemical_list: D000998:Antiviral Agents; D004279:DNA, Viral; C109367:UL54 protein, Human herpesvirus 5; D014764:Viral Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); C075365:ganciclovir kinase; D004259:DNA-Directed DNA Polymerase; D015774:Ganciclovir
keywords: Cytomegalovirus; HCMV; Hematopoietic stem cell transplant; Resistance mutations; UL54; UL97