南方医科大学学报Issue(4)：519-522,4.DOI:10.3969/j.issn.1673-4254.2014.04.17
造血干细胞移植后巨细胞病毒药敏检测及UL97基因突变分析
Drug susceptibility and UL97 gene mutation analysis of cytomegalovirus in recipients of hemato-poietic stem cell transplantation
摘要
Abstract
Objective To monitor human cytomegalovirus (HCMV) drug resistance in recipients of hematopoietic stem cell transplantation by phenotypic and genotypic methods. Methods HCMV clinical isolates was isolated from the urine of hematopoietic stem cell transplantation recipients treated with GCV. Tissue cell infection median dose (TCID50) of the isolates was calculated using Reed-Muench method, and their drug susceptibility was determined by plaque reduction assay. We amplified the UL97 DNA fragment of the virus by nested PCR followed by automated DNA sequencing. Results HCMV clinical strain isolated from the urine samples of the recipients using a human fibroblast cell line showed a TCID50 value of 10-4.618/0.1ml and a 50%inhibitory concentration (IC50) to GCV of 5.847 µmol/L, suggesting its sensitivity to GCV. Alignment with the AD169 DNA reference sequence identified 4 point mutations of the virus at 1509 (T-C), 1575 (C-T), 1794 (T-C), and 1815 (C-G), and only the last mutation resulted in one amino acid mutation to D605E. No gene mutation was found in relation to GCV resistance. Conclusions Phenotypic and genotypic assays were established to examine antiviral drug resistance of HCMV in recipients of hematopoietic stem cell transplantation. We did not find any drug resistance of the clinical HCMV isolate.关键词
人巨细胞病毒/耐药/丙氧鸟苷/基因突变Key words
cytomegalovirus/resistance/ganciclovir/mutation引用本文复制引用
曾爱红,董德坤,崔喜梅,郭明明,邹德志,方建培..造血干细胞移植后巨细胞病毒药敏检测及UL97基因突变分析[J].南方医科大学学报,2014,(4):519-522,4.基金项目
广东省科技计划项目 （）
