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

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-resol...

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Bibliographic Details
Published in:Archives of virology 2012-03, Vol.157 (3), p.475-481
Main Authors: Zhao, Xiao-Tao, Zhou, Dan-Qiu, Wu, Shuai, Chen, Yue-Wen, Shao, Yong, Zhang, Jie, Xia, Chang-Sheng, Wang, Ke-Peng, Yang, Hong, Wan, Jun, Yu, Bo, Zhang, Zheng, Zhang, Wei
Format: Article
Language:English
Subjects:
Antiviral Agents - pharmacology
Biological and medical sciences
Biomedical and Life Sciences
Biomedicine
Cloning
Cytomegalovirus
Cytomegalovirus - drug effects
Cytomegalovirus - genetics
Dermatology
DNA, Viral - genetics
Drug Resistance, Viral
Fundamental and applied biological sciences. Psychology
Ganciclovir - pharmacology
Genotype
Human cytomegalovirus
Humans
Infectious Diseases
Kinases
Medical Microbiology
Microbiology
Miscellaneous
Molecular Typing
Mutation
Mutation, Missense
Original Article
Pathogens
Phosphotransferases (Alcohol Group Acceptor) - genetics
Sensitivity and Specificity
Transition Temperature
Virology
Virology - methods
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Summary: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.
ISSN:0304-8608
1432-8798
DOI:10.1007/s00705-011-1173-y