title: Detection of Two Drug-Resistance Mutants of the Cytomegalovirus by High-Resolution Melting Analysis.
abstract: BACKGROUND
  Human cytomegalovirus (CMV) is an opportunistic pathogen that can be treated with ganciclovir. Mutations in the UL97 gene of CMV render the virus ganciclovir resistance. These include H520Q and C603W mutations, against which we developed a novel genotyping assay for their identification.
  
  
  METHODS
  PCR reactions were performed to amplify fragments of the UL97 gene containing H520Q or C603W mutations. High resolution melting analysis (HRMA) coupled with unlabeled DNA probes was employed to identify the shift in melting temperature of the probe-template complex, which reflexes the presence of point mutations.
  
  
  RESULTS
  Melting point analysis performed on the dimeric DNA of PCR products of UL97 gene could not identify mutations in the gene. When coupled to unlabeled probes, point mutations in UL97 can be identified by analyzing the melting curve of probe-template complex. When WT and mutant UL97 DNAs were mixed together to mimic heterogeneous viral population in clinical samples, the genotyping assay is sensitive enough to detect H520Q and C603W mutants that constitute 10% of total DNA input.
  
  
  CONCLUSIONS
  Probe-based HRMA is effective in detecting H520Q and C603W mutations in the UL97 gene of CMV.
authors.affiliation: Biomedical Research Institute, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, Guangdong, China.|Department of Dermatology, Shenzhen Hospital Peking University, Shenzhen, Guangdong, China.|Shenzhen Key Lab for Translational Medicine of Dermatology, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, Guangdong, China.
authors.forename: Xiao-Fan
authors.initials: XF
authors.lastname: Chen
authors.affiliation: Department of Intervention and Vascular Surgery, Peking University Third Hospital, Beijing, China.
authors.forename: Tian-Run
authors.initials: TR
authors.lastname: Li
authors.affiliation: Department of Clinical Laboratory, Shenzhen Hospital Peking University, Shenzhen, Guangdong, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Yang
authors.affiliation: Department of Dermatology, Shenzhen Hospital Peking University, Shenzhen, Guangdong, China.|Shenzhen Key Lab for Translational Medicine of Dermatology, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, Guangdong, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Shao
authors.affiliation: Department of Dermatology, Shenzhen Hospital Peking University, Shenzhen, Guangdong, China.|Shenzhen Key Lab for Translational Medicine of Dermatology, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, Guangdong, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Biomedical Research Institute, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, Guangdong, China.|Shenzhen Key Lab for Translational Medicine of Dermatology, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, Guangdong, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Department of Dermatology, Shenzhen Hospital Peking University, Shenzhen, Guangdong, China.|Shenzhen Key Lab for Translational Medicine of Dermatology, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, Guangdong, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Yu
authors.affiliation: Biomedical Research Institute, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, Guangdong, China.|Shenzhen Key Lab for Translational Medicine of Dermatology, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, Guangdong, China.
authors.forename: Zhun
authors.initials: Z
authors.lastname: Wei
authors.affiliation: Department of Dermatology, Shenzhen Hospital Peking University, Shenzhen, Guangdong, China.|Shenzhen Key Lab for Translational Medicine of Dermatology, Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center, Shenzhen, Guangdong, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Wu
authors.affiliation: Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital, Shenzhen, Guangdong, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Yu
mesh_terms: D003587:Cytomegalovirus / Q000235:genetics*; D015342:DNA Probes / Q000378:metabolism; D024882:Drug Resistance, Viral / Q000235:genetics*; D006801:Humans; D009154:Mutation / Q000235:genetics*; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D015342:DNA Probes
keywords: C603W; H520Q; UL97; cytomegalovirus; high resolution melting analysis