Drug susceptibility and UL97 gene mutation analysis of cytomegalovirus in recipients of hemato-poietic stem cell transplantation
10.3969/j.issn.1673-4254.2014.04.17
- VernacularTitle: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
- Author:
Aihong ZENG
1
;
Dekun DONG
;
Ximei CUI
;
Mingming GUO
;
Dezhi ZOU
;
Jianpei FANG
Author Information
1. 中山大学附属一院急诊科
- Keywords:
cytomegalovirus;
resistance;
ganciclovir;
mutation
- From:
Journal of Southern Medical University
2014;(4):519-522
- CountryChina
- Language:Chinese
-
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.
