Antiviral drug resistance in human cytomegalovirus

Chou, Sunwen

doi:10.1034/j.1399-3062.1999.010204.x

Cited by 85 publications

(101 citation statements)

References 49 publications

(1 reference statement)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The high level of viral replication (due to primary infection), a prolonged antiviral therapy, and administration of immunosuppressive drugs such as cyclosporine and corticosteroids as well as the conditioning regimen required for HSCT (and therefore an incompetent immune response with low CD4 and CD8 T-cell counts) are all factors that favor the emergence of resistant viruses during a course of therapy (2).…”

mentioning

confidence: 99%

Fatal Reactivation of Postnatal Cytomegalovirus Infection with Rapid Emergence of Ganciclovir Resistance in an Infant after Allogeneic Stem Cell Transplantation

et al. 2005

View full text Add to dashboard Cite

Human cytomegalovirus (HCMV) can cause serious problems after hematopoietic stem cell transplantation. The death of a pediatric transplant recipient after reactivation of a postnatal HCMV infection with bilateral retinitis and pneumonitis is described. Sequencing of the HCMV UL97 region revealed a compartment-specific mutation (H520Q) in urine conferring ganciclovir resistance. CASE REPORTA 3-month-old infant had been diagnosed with juvenile myelomonocytic leukemia (JMML), a rare myeloproliferative disorder of childhood, and was admitted 2 months later for hematopoietic stem cell transplantation (HSCT). An HLAcompatible (HLA-A, -B, -C, -DRB1, and -DQB1-matched, one mismatch in HLA-DPB1), unrelated, human cytomegalovirus (HCMV)-seronegative donor was identified, and the infant was scheduled for an allogeneic HSCT at the age of 6 months. According to the European Working Group on Myelodysplastic Syndrome protocol, the conditioning regimen included busulfan, cyclophosphamide, and melphalan. Antithymocyte globulin was administered as prophylaxis for graftversus-host disease (GVHD) prior to transplantation. Posttransplantation, cyclosporine was used for GVHD prophylaxis, and only mild acute GVHD confined to the skin (maximum, 25%) had occurred. Hematopoietic reconstitution was initially protracted. Analysis of blood DNA short terminal repeats revealed 100% donor chimerism. At the time of pretransplantation evaluation, the HCMV-specific immunoglobulin G (IgG) assay was weakly positive whereas HCMV-specific IgM and HCMV DNA assays were negative. The infant had been breast-fed, but breast milk had not been tested for HCMV DNA by PCR. A retrospective PCR analysis from blood of the Guthrie card obtained during the first days of life of the newborn demonstrated HCMV DNA negativity. Therefore, HCMV-specific IgG antibodies were considered to be of maternal origin, for the mother was HCMV seropositive.Due to severe thrombocytopenia, the infant received a leukocyte-depleted platelet transfusion from an HCMV-seropositive donor at the age of 5 months. One week later, HCMV IgM was detected with increasing sample-to-cutoff ratios from 2.8 to 6.4 over the next 3 weeks. However, during this time, HCMV DNA PCR from leukocytes and urine was negative, and HSCT was performed. Acyclovir prophylaxis was routinely started 1 day after transplantation.On day 10 posttransplantation systemic active HCMV infection was first detected by PCR from blood. Seven days later anti-HCMV therapy was started with intravenous ganciclovir (GCV) at a dose of 2 ϫ 5 mg/kg of body weight per day (Fig. 1). Due to fluctuating HCMV loads and pp65-positive cells in the antigenemia assay as well as increasing myelosuppression, therapy was briefly switched to foscarnet (PFA), followed by alternate medication of GCV and PFA. However, response to therapy with GCV remained poor, which was reflected in continuing fluctuating levels of HCMV antigenemia and the development of HCMV retinitis. On day ϩ62, the infant developed fever with increasing viral loads of HCMV and Ep...

show abstract

mentioning

confidence: 99%

Fatal Reactivation of Postnatal Cytomegalovirus Infection with Rapid Emergence of Ganciclovir Resistance in an Infant after Allogeneic Stem Cell Transplantation

et al. 2005

View full text Add to dashboard Cite

show abstract

“…The in vitro assay determines the antiviral's ability to reduce the number of lytic plaques, which quantifies the cytopathic effect induced by the virus. This can be determined as a 50% inhibitory concentration (IC 50 ) (Chou et al, 1999;Landry et al, 2000). This is a difficult method that requires trained personnel and standard antiviral concentrations.…”

Section: Laboratory Methods For the Detection Of Antiviral Drugsmentioning

confidence: 99%

Point Mutations and Antiviral Drug Resistance

Arellano‐Galindo¹,

Barrón²,

Vargas-Infante³

et al. 2012

Point Mutation

View full text Add to dashboard Cite

“…All experiments were performed three times independently. GCV resistance was defined as cells with an IC 50 exceeding 6 M to 12 M GCV (6,7,14).…”

Section: Cells and Virusmentioning

confidence: 99%

Use of Time-Saving Flow Cytometry for Rapid Determination of Resistance of Human Cytomegalovirus to Ganciclovir

Lee

Choi

et al. 2005

J Clin Microbiol

View full text Add to dashboard Cite

There are two ways to assess the susceptibility of human cytomegalovirus (HCMV) to ganciclovir (GCV): one is a genotypic test that detects resistance-related mutations and the other is a phenotypic test that actually assesses susceptibility. The advantages of genotyping the UL97 gene are its rapidity and accuracy. However, to detect novel mutations or mutations affecting the UL54 DNA polymerase, a phenotypic test such as the plaque reduction assay (PRA) is also required. To avoid the shortcomings of PRA such as its time-consuming nature and labor-intensiveness, we developed a time-saving fluorescence-activated cell sorting (TS-FACS) technique. We obtained a GCV 50% inhibitory concentration (IC 50 ) from five clinical isolates and an HCMV laboratory strain (AD169) and compared the results with those from the PRA. The laboratory strain and three clinical isolates were sensitive to GCV. Although there was a minor discrepancy in the case of one of the three isolates, the GCV IC 50 values obtained by TS-FACS analysis correlated well with the results of the PRA. The remaining two isolates were resistant to GCV; one was GCV resistant due to the mutation M460V, and the GCV IC 50 results obtained by TS-FACS analysis and by PRA were also comparable. The advantages of TS-FACS analysis are the shorter time required, the possibility of automation, and its comparability to PRA, considered the gold standard. Thus, TS-FACS analysis may be useful as an alternative to PRA in the clinic.Human cytomegalovirus (HCMV) is a betaherpesvirus that causes serious diseases in immunocompromised hosts, particularly transplantation recipients. Drugs used currently against HCMV include ganciclovir (GCV), foscarnet, and cidofovir, and of these, GCV is the initial drug of choice (11, 13). However, the advent of HCMV resistant to GCV has made control of HCMV difficult. Mutant HCMV strains strongly resistant to GCV are unable to phosphorylate GCV owing to mutations in key regions of the UL97 gene (domains VIII, VI, and IX) (1,2,5,8,9,21,26). Mutations in UL54, the viral DNA polymerase, also confer resistance to GCV (10,15,24).There are two types of methods for assessing the susceptibility of HCMV to GCV: one is genotypic and detects the resistance-related mutant genes, and the other is phenotypic and assesses resistance directly (3, 13). The genotypic methods include DNA sequencing, detection of restriction fragment length polymorphisms (RFLP), and probe-specific and primerspecific hybridization. DNA sequencing is the reference method for detecting resistance-related GCV mutations, and RFLP has been widely used to detect GCV resistance due to mutations in the UL97 gene (4, 7). Although genotypic tests are fast, phenotypic tests are necessary to detect novel mutations and mutations in the UL54 DNA polymerase gene. Examples of phenotypic tests are the plaque reduction assay (PRA), in situ enzyme-linked immunosorbent assay, DNA reduction assay, and flow cytometry-based assay. The flow cytometric assay counts trypsinized fibroblasts expressing the HCM...

show abstract

Antiviral drug resistance in human cytomegalovirus

Cited by 85 publications

References 49 publications

Fatal Reactivation of Postnatal Cytomegalovirus Infection with Rapid Emergence of Ganciclovir Resistance in an Infant after Allogeneic Stem Cell Transplantation

Fatal Reactivation of Postnatal Cytomegalovirus Infection with Rapid Emergence of Ganciclovir Resistance in an Infant after Allogeneic Stem Cell Transplantation

Point Mutations and Antiviral Drug Resistance

Use of Time-Saving Flow Cytometry for Rapid Determination of Resistance of Human Cytomegalovirus to Ganciclovir

Contact Info

Product

Resources

About