Phenotyping of cytomegalovirus drug resistance mutations by using recombinant viruses incorporating a reporter gene.

Abstract:

:A new recombinant phenotyping method was developed for the analysis of drug resistance mutations in human cytomegalovirus (CMV). CMV strain T2211 was derived from strain AD169 by inserting unique restriction sites and a secreted alkaline phosphatase (SEAP) reporter gene for rapid viral quantitation. Specific viral UL97 and pol gene mutations were transferred by recombination into T2211, and their drug resistance phenotypes (for ganciclovir, foscarnet, or cidofovir) were determined by the drug concentrations required to reduce supernatant SEAP activity by 50% (IC50). Changes in the IC50 conferred by the mutations tested (UL97 M460V, C592G, A594V, and L595S and pol del981-2) were similar to those previously reported in marker transfer and conventional plaque reduction assays. The combination of UL97 C592G and pol del981-2 conferred much higher ganciclovir resistance than either mutation alone. The UL97 polymorphism D605E had no measurable effect on ganciclovir susceptibility, alone or in combination with common UL97 resistance mutations. Transfer into strain T2211 facilitates the phenotyping of newly observed mutations, combinations of mutations, and clinical CMV sequences without an accompanying viral isolate.

authors

Chou S,Van Wechel LC,Lichy HM,Marousek GI

doi

10.1128/AAC.49.7.2710-2715.2005

subject

Has Abstract

pub_date

2005-07-01 00:00:00

pages

2710-5

issue

7

eissn

0066-4804

issn

1098-6596

pii

49/7/2710

journal_volume

49

pub_type

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