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RAPID DETECTION OF CYTOMEGALOVIRUS STRAINS RESISTANT TO GANCICLOVIR THROUGH MUTATIONS WITHIN THE GENE UL97
被引:19
作者
:
ALAIN, S
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV PARIS 07, HOP LARIBOISIERE, SERV BACTERIOL VIROL, F-75010 PARIS, FRANCE
ALAIN, S
MAZERON, MC
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV PARIS 07, HOP LARIBOISIERE, SERV BACTERIOL VIROL, F-75010 PARIS, FRANCE
MAZERON, MC
PEPIN, JM
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV PARIS 07, HOP LARIBOISIERE, SERV BACTERIOL VIROL, F-75010 PARIS, FRANCE
PEPIN, JM
MORINET, F
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV PARIS 07, HOP LARIBOISIERE, SERV BACTERIOL VIROL, F-75010 PARIS, FRANCE
MORINET, F
RASKINE, L
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV PARIS 07, HOP LARIBOISIERE, SERV BACTERIOL VIROL, F-75010 PARIS, FRANCE
RASKINE, L
SANSONLEPORS, MJ
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV PARIS 07, HOP LARIBOISIERE, SERV BACTERIOL VIROL, F-75010 PARIS, FRANCE
SANSONLEPORS, MJ
机构
:
[1]
UNIV PARIS 07, HOP LARIBOISIERE, SERV BACTERIOL VIROL, F-75010 PARIS, FRANCE
[2]
HOP BICHAT CLAUDE BERNARD, SERV VIROL, PARIS, FRANCE
[3]
HOP ST LOUIS, SERV BACTERIOL VIROL, PARIS, FRANCE
来源
:
MOLECULAR AND CELLULAR PROBES
|
1993年
/ 7卷
/ 06期
关键词
:
HUMAN CYTOMEGALOVIRUS;
ANTIVIRAL DRUG;
KINASE;
ADN POLYMERASE;
GANCICLOVIR RESISTANCE;
GENE UL97;
POINT MUTATION;
DELETION;
D O I
:
10.1006/mcpr.1993.1072
中图分类号
:
Q5 [生物化学];
学科分类号
:
071010 ;
081704 ;
摘要
:
The human cytomegalovirus (HCMV) gene UL97 product was shown to play an important role in phosphorylation of ganciclovir (GCV) in HCMV-infected cells. The deletion of the 4 amino-acid sequence AACR confers resistance to a laboratory mutant. The aim of this study was to develop a rapid and simple method to detect mutations within the 12 base pair (bp) fragment encoding AACR, from isolates and clinical specimens (urine, bronchoalveolar lavage, cerebral spinal fluid samples). A target region encompassing this 12 bp sequence was amplified by a single-step PCR assay from HCMV isolates and reference strains, and a two-step procedure from clinical specimens. Reaction products were submitted to restriction enzyme analysis and dot-blot hybridization assay. Two biotinylated probes were used: one probe (DL) overlapping the 12 bp region; and a control probe with similar length and GC content. Hybridization was performed under conditions allowing the detection of one bp deletion (HCMV strain susceptibility to GCV was determined by a rapid late antigen synthesis reduction assay.) The control probe hybridized to the UL97 sequence amplified from all 23 tested isolates and the reference strains. The DL probe gave a positive signal with GCV-susceptible strains; no signal was obtained for five out of seven resistant isolates, and for a laboratory mutant derived from the strain AD169. Restriction analysis of amplification products showed different patterns suggesting this region can be involved in various DNA changes. The results from assays directly performed on 10 clinical samples correlated with GCV strain susceptibility determination. This rapid and simple method for detecting mutations in this region of the gene UL97 could be a useful tool for identification of some GCV-resistant strains from isolates and directly from clinical specimens. © 1993 Academic Press. All rights reserved.
引用
收藏
页码:487 / 495
页数:9
相关论文
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←
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2
3
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共 24 条
[1]
A HUMAN CYTOMEGALOVIRUS MUTANT RESISTANT TO THE NUCLEOSIDE ANALOG 9-([2-HYDROXY-1-(HYDROXYMETHYL)ETHOXY]METHYL)GUANINE (BW B759U) INDUCES REDUCED LEVELS OF BW B759U TRIPHOSPHATE
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0
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0
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WELLCOME RES LABS,DEPT EXPTL THERAPY,RES TRIANGLE PK,NC 27709
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0
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METABOLIC-ACTIVATION OF THE NUCLEOSIDE ANALOG 9-([2-HYDROXY-L-(HYDROXYMETHYL)ETHOXY]METHYL)GUANINE IN HUMAN-DIPLOID FIBROBLASTS INFECTED WITH HUMAN CYTOMEGALO-VIRUS
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