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学科主题: 临床医学
题名:
Development of SYBR Green I-based real-time PCR assay for detection of drug resistance mutations in cytomegalovirus
作者: Liu, Jing-bo; Zhang, Zheng
关键词: cytomegalovirus ; real-time PCR ; SYBR green ; ganciclovir
刊名: JOURNAL OF VIROLOGICAL METHODS
发表日期: 2008-04-01
DOI: 10.1016/j.jviromet.2007.12.011
卷: 149, 期:1, 页:129-135
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology
研究领域[WOS]: Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Virology
关键词[WOS]: CONFERRING GANCICLOVIR RESISTANCE ; STEM-CELL TRANSPLANTATION ; ANTIVIRAL DRUGS ; RAPID DETECTION ; UL97 GENE ; INFECTION ; CODON-460 ; CHILDREN
英文摘要:

Ganciclovir (GCV) is an antiviral drug that is used to treat cytomegalovirus (CMV) infection. However, long-term monotherapy does not commonly result in complete suppression of viral replication and is associated with the emergence of resistant mutants. In this study, a method for detecting CMV resistance mutations was carried out by real-time amplification refractory mutation system PCR (real-time ARMS PCR) using SYBR Green I fluorescent dye. Three recombinant plasmids were constructed by overlapping extension PCR to be used as standard mutation or wild-type models. Four pairs of primers were used to amplify the approximately 150 bp of the UL97 gene spanning codon 460, where mutations associated with resistance to GCV invariably occur. As little as 20% mutants DNA in 10(7) copies/ml mixture DNA were detected. Though this approach was not more sensitive than PCR-restriction fragment length polymorphism (RFLP) for the detection of the presence of mixtures, it was a high-throughput and automation method, and the specific mutation type can be deduced by the real-time ARMS PCR data. Overall, this study has demonstrated an approach that could be a sensitive and rapid method for the detection of GCV resistance-associated mutation in CMV. (C) 2008 Elsevier B.V. All rights reserved.

语种: 英语
WOS记录号: WOS:000255300700019
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/63808
Appears in Collections:北京大学第二临床医学院_期刊论文

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作者单位: Peking Univ, Peoples Hosp, Dept Clin Lab, Beijing 100044, Peoples R China

Recommended Citation:
Liu, Jing-bo,Zhang, Zheng. Development of SYBR Green I-based real-time PCR assay for detection of drug resistance mutations in cytomegalovirus[J]. JOURNAL OF VIROLOGICAL METHODS,2008,149(1):129-135.
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