title: [Drug susceptibility and UL97 gene mutation analysis of cytomegalovirus in recipients of hematopoietic stem cell transplantation].
abstract: OBJECTIVE
  To monitor human cytomegalovirus (HCMV) drug resistance in recipients of hematopoietic stem cell transplantation by phenotypic and genotypic methods.
  
  
  METHODS
  HCMV clinical isolates was isolated from the urine of hematopoietic stem cell transplantation recipients treated with GCV. Tissue cell infection median dose (TCID50) of the isolates was calculated using Reed-Muench method, and their drug susceptibility was determined by plaque reduction assay. We amplified the UL97 DNA fragment of the virus by nested PCR followed by automated DNA sequencing.
  
  
  RESULTS
  HCMV clinical strain isolated from the urine samples of the recipients using a human fibroblast cell line showed a TCID50 value of 10(-4.618)/0.1 ml and a 50% inhibitory concentration (IC50) to GCV of 5.847 µmol/L, suggesting its sensitivity to GCV. Alignment with the AD169 DNA reference sequence identified 4 point mutations of the virus at 1509 (T-C), 1575 (C-T), 1794 (T-C), and 1815 (C-G), and only the last mutation resulted in one amino acid mutation to D605E. No gene mutation was found in relation to GCV resistance.
  
  
  CONCLUSIONS
  Phenotypic and genotypic assays were established to examine antiviral drug resistance of HCMV in recipients of hematopoietic stem cell transplantation. We did not find any drug resistance of the clinical HCMV isolate.
authors.affiliation: Department of Emergency Medicine, First Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510080, China.E-mail: Janezah @163.com.
authors.forename: Aihong
authors.initials: A
authors.lastname: Zeng
authors.forename: Dekun
authors.initials: D
authors.lastname: Dong
authors.forename: Ximei
authors.initials: X
authors.lastname: Cui
authors.forename: Mingming
authors.initials: M
authors.lastname: Guo
authors.forename: Dezhi
authors.initials: D
authors.lastname: Zou
authors.forename: Jianpei
authors.initials: J
authors.lastname: Fang
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology; D002460:Cell Line; D003587:Cytomegalovirus / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D024882:Drug Resistance, Viral / Q000235:genetics*; D015774:Ganciclovir / Q000494:pharmacology; D005814:Genes, Viral; D005838:Genotype; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D009154:Mutation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics*
chemical_list: D000998:Antiviral Agents; D017853:Phosphotransferases (Alcohol Group Acceptor); C075365:ganciclovir kinase; D015774:Ganciclovir