Recombinant phenotyping of cytomegalovirus sequence variants detected after 200 or 100 days of valganciclovir prophylaxis

Sunwen Chou, Gail Marousek, Guy Boivin, Nathalie Goyette, Mahdi Farhan, Jane A L Ives, Robert Elston

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Background. In a phase III controlled trial IMproved Protection Against Cytomegalovirus in Transplantation (IMPACT) comparing 200 with 100 days of valganciclovir prophylaxis in 318 cytomegalovirus D+/R- kidney transplant recipients, an equal number of patients (n=3 per arm) had known ganciclovir resistance mutations detected during viral breakthrough. In addition, many other viral sequence variants were observed that were of unknown significance for ganciclovir resistance. Recombinant phenotyping was performed to determine whether the previously uncharacterized genotypic changes affected ganciclovir susceptibility, especially in those receiving the longer duration of prophylaxis. Methods. Sequences encoding individual amino acid substitutions in the UL97 kinase or UL54 DNA polymerase gene were transferred by recombination into a cloned cytomegalovirus laboratory strain, followed by reporter-based yield reduction phenotypic assay of the resulting virus for ganciclovir susceptibility. Results. Twenty-six uncharacterized amino acid substitutions were detected, 2 in UL97 and 24 in UL54. All 10 substitutions in the 200-day arm and 9 of 17 substitutions in the 100-day arm (prioritized based on location and conservation) were selected for phenotyping; one substitution was detected in both subsets. Results were generated for nine of ten 200-day and eight of nine 100-day substitutions, with no substitution demonstrating a significant reduction in ganciclovir susceptibility. The two remaining amino acid substitutions, both in UL54, were not evaluated because of poor viral viability. Conclusion. Phenotypic evaluation of previously uncharacterized viral genotypes in the 200-day valganciclovir prophylaxis group showed no evidence of an increased incidence of genotypic ganciclovir resistance when compared with those in the 100-day prophylaxis group.

Original languageEnglish (US)
Pages (from-to)1409-1413
Number of pages5
JournalTransplantation
Volume90
Issue number12
DOIs
StatePublished - Dec 27 2010

Fingerprint

Ganciclovir
Cytomegalovirus
Amino Acid Substitution
DNA-Directed DNA Polymerase
Genetic Recombination
valganciclovir
Phosphotransferases
Transplantation
Genotype
Viruses
Kidney
Mutation
Incidence
Genes

Keywords

  • Cytomegalovirus
  • Prophylaxis
  • Resistance
  • Transplant
  • Valganciclovir

ASJC Scopus subject areas

  • Transplantation

Cite this

Recombinant phenotyping of cytomegalovirus sequence variants detected after 200 or 100 days of valganciclovir prophylaxis. / Chou, Sunwen; Marousek, Gail; Boivin, Guy; Goyette, Nathalie; Farhan, Mahdi; Ives, Jane A L; Elston, Robert.

In: Transplantation, Vol. 90, No. 12, 27.12.2010, p. 1409-1413.

Research output: Contribution to journalArticle

Chou, Sunwen ; Marousek, Gail ; Boivin, Guy ; Goyette, Nathalie ; Farhan, Mahdi ; Ives, Jane A L ; Elston, Robert. / Recombinant phenotyping of cytomegalovirus sequence variants detected after 200 or 100 days of valganciclovir prophylaxis. In: Transplantation. 2010 ; Vol. 90, No. 12. pp. 1409-1413.
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AU - Chou, Sunwen

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AU - Boivin, Guy

AU - Goyette, Nathalie

AU - Farhan, Mahdi

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AB - Background. In a phase III controlled trial IMproved Protection Against Cytomegalovirus in Transplantation (IMPACT) comparing 200 with 100 days of valganciclovir prophylaxis in 318 cytomegalovirus D+/R- kidney transplant recipients, an equal number of patients (n=3 per arm) had known ganciclovir resistance mutations detected during viral breakthrough. In addition, many other viral sequence variants were observed that were of unknown significance for ganciclovir resistance. Recombinant phenotyping was performed to determine whether the previously uncharacterized genotypic changes affected ganciclovir susceptibility, especially in those receiving the longer duration of prophylaxis. Methods. Sequences encoding individual amino acid substitutions in the UL97 kinase or UL54 DNA polymerase gene were transferred by recombination into a cloned cytomegalovirus laboratory strain, followed by reporter-based yield reduction phenotypic assay of the resulting virus for ganciclovir susceptibility. Results. Twenty-six uncharacterized amino acid substitutions were detected, 2 in UL97 and 24 in UL54. All 10 substitutions in the 200-day arm and 9 of 17 substitutions in the 100-day arm (prioritized based on location and conservation) were selected for phenotyping; one substitution was detected in both subsets. Results were generated for nine of ten 200-day and eight of nine 100-day substitutions, with no substitution demonstrating a significant reduction in ganciclovir susceptibility. The two remaining amino acid substitutions, both in UL54, were not evaluated because of poor viral viability. Conclusion. Phenotypic evaluation of previously uncharacterized viral genotypes in the 200-day valganciclovir prophylaxis group showed no evidence of an increased incidence of genotypic ganciclovir resistance when compared with those in the 100-day prophylaxis group.

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