Archives of Virology

, Volume 157, Issue 3, pp 475–481

Genotyping cytomegalovirus UL97 mutations by high-resolution melting analysis with unlabeled probe

Authors

  • Xiao-Tao Zhao
    • Department of Clinical LaboratoryPeking University People’s Hospital
  • Dan-Qiu Zhou
    • Department of Laboratory MedicineJinshan Hospital, Fudan University
  • Shuai Wu
    • College of Pharmacy, Ji-Nan University
  • Yue-Wen Chen
    • College of Pharmacy, Ji-Nan University
  • Yong Shao
    • Shenzhen Key Laboratory for Translational Medicine of DermatologyShenzhen-PKU-HKUST Medical Center
    • Biomedical Research Institute, Shenzhen-PKU-HKUST Medical Center
    • Department of DermatologyShenzhen Hospital Peking University
  • Jie Zhang
    • Shenzhen Key Laboratory for Translational Medicine of DermatologyShenzhen-PKU-HKUST Medical Center
    • Biomedical Research Institute, Shenzhen-PKU-HKUST Medical Center
    • Department of DermatologyShenzhen Hospital Peking University
  • Chang-Sheng Xia
    • Department of Clinical LaboratoryPeking University People’s Hospital
  • Ke-Peng Wang
    • Biomedical Research Institute, Shenzhen-PKU-HKUST Medical Center
  • Hong Yang
    • Shenzhen Key Laboratory for Translational Medicine of DermatologyShenzhen-PKU-HKUST Medical Center
    • Biomedical Research Institute, Shenzhen-PKU-HKUST Medical Center
  • Jun Wan
    • Shenzhen Key Laboratory for Translational Medicine of DermatologyShenzhen-PKU-HKUST Medical Center
    • Biomedical Research Institute, Shenzhen-PKU-HKUST Medical Center
    • Division of Life ScienceThe Hong Kong University of Science and Technology
    • Shenzhen Key Laboratory for Translational Medicine of DermatologyShenzhen-PKU-HKUST Medical Center
    • Biomedical Research Institute, Shenzhen-PKU-HKUST Medical Center
    • Department of DermatologyShenzhen Hospital Peking University
    • Department of Clinical LaboratoryPeking University People’s Hospital
    • College of Pharmacy, Ji-Nan University
    • Shenzhen Key Laboratory for Translational Medicine of DermatologyShenzhen-PKU-HKUST Medical Center
    • Biomedical Research Institute, Shenzhen-PKU-HKUST Medical Center
Original Article

DOI: 10.1007/s00705-011-1173-y

Cite this article as:
Zhao, X., Zhou, D., Wu, S. et al. Arch Virol (2012) 157: 475. doi:10.1007/s00705-011-1173-y
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Abstract

Human cytomegalovirus (CMV) is an opportunistic pathogen, and infections with this virus can be treated with ganciclovir (GCV). Most GCV-resistant clinical CMV isolates contain a mutation in the UL97 gene. Genotypic assays for diagnostic screening of GCV-resistant CMV have been developed. High-resolution melting analysis (HRMA) with unlabeled probe is considered a perfect tool for this purpose. In this study, we have developed an HRMA-based genotypic test for the detection of UL97 mutations. Wild type and M460V/I mutants of UL97 were constructed. HRMA with unlabeled probe was used as a genotyping method for the detection of M460V/I mutations. The melting peaks obtained directly from PCR products did not enable us to distinguish the wild type from M460 mutants. The sensitivity and accuracy of HRMA were dramatically improved by using unlabeled probe. HRMA with unlabeled probe successfully distinguished M460V from M460I and served well for the detection of M460V/I mutations in clinical samples. HRMA with unlabeled probe proves to be a sensitive and cost-effective genotyping method for the detection of M460 mutations.

Copyright information

© Springer-Verlag 2011