Archives of Virology

, Volume 157, Issue 3, pp 475–481 | Cite as

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

  • Xiao-Tao Zhao
  • Dan-Qiu Zhou
  • Shuai Wu
  • Yue-Wen Chen
  • Yong Shao
  • Jie Zhang
  • Chang-Sheng Xia
  • Ke-Peng Wang
  • Hong Yang
  • Jun Wan
  • Bo Yu
  • Zheng Zhang
  • Wei Zhang
Original Article

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.

Notes

Acknowledgments

The study was supported by research grants from the Shenzhen Science and Technology Project. We thank Shenzhen Biomedical Research Support Platform for technical help.

Conflict of interest

None.

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Xiao-Tao Zhao
    • 1
  • Dan-Qiu Zhou
    • 2
  • Shuai Wu
    • 3
  • Yue-Wen Chen
    • 3
  • Yong Shao
    • 4
    • 5
    • 6
  • Jie Zhang
    • 4
    • 5
    • 6
  • Chang-Sheng Xia
    • 1
  • Ke-Peng Wang
    • 5
  • Hong Yang
    • 4
    • 5
  • Jun Wan
    • 4
    • 5
    • 7
  • Bo Yu
    • 4
    • 5
    • 6
  • Zheng Zhang
    • 1
  • Wei Zhang
    • 3
    • 4
    • 5
  1. 1.Department of Clinical LaboratoryPeking University People’s HospitalPekingChina
  2. 2.Department of Laboratory MedicineJinshan Hospital, Fudan UniversityShanghaiChina
  3. 3.College of Pharmacy, Ji-Nan UniversityGuangdongChina
  4. 4.Shenzhen Key Laboratory for Translational Medicine of DermatologyShenzhen-PKU-HKUST Medical CenterShenzhenChina
  5. 5.Biomedical Research Institute, Shenzhen-PKU-HKUST Medical CenterShenzhenChina
  6. 6.Department of DermatologyShenzhen Hospital Peking UniversityShenzhenChina
  7. 7.Division of Life ScienceThe Hong Kong University of Science and TechnologyHong KongChina

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