Detection of ganciclovir resistance mutations by pyrosequencing in HCMV-infected pediatric patients
| Autor: | Irene Vanni, Giuseppe Morreale, C. Cirillo, Fabio Benzi, Giulia Cassina, Mauro S. Malnati, Eddi Di Marco, Giovanni Melioli, Elisabetta Ugolotti, Roberto Biassoni, Emilio Cristina |
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| Rok vydání: | 2012 |
| Předmět: |
Ganciclovir
Human cytomegalovirus Viral protein viruses Molecular Sequence Data Mutant Mutation Missense Cytomegalovirus Microbial Sensitivity Tests Biology medicine.disease_cause Antiviral Agents Virology medicine SNPs analysis Humans Viral Gene HCMV Mutation Drug-resistance-associated mutation Pyrosequencing QPCR UL97 mutations Cytomegalovirus Infections DNA Viral Infant Infant Newborn Phosphotransferases (Alcohol Group Acceptor) Sequence Analysis DNA DNA Newborn Resistance mutation medicine.disease Phenotype Infectious Diseases Missense Sequence Analysis medicine.drug |
| Zdroj: | Journal of Clinical Virology. 54:48-55 |
| ISSN: | 1386-6532 |
| DOI: | 10.1016/j.jcv.2012.01.006 |
| Popis: | Background Human cytomegalovirus (HCMV) is an opportunistic pathogen especially for immuno-suppressed subjects that might develop pharmacological resistance in patients undergoing prolonged antiviral treatment. Ganciclovir (GCV) is the drug used as first choice therapy in affected children and a GCV-resistant phenotype is mainly linked to mutations of the viral protein kinase UL97. Objectives Here a new quantitative pyrosequence (PSQ) method is presented that allows detection and quantification of the viral species carrying the more frequent UL97 mutations responsible for GCV resistance in clinical samples (>80% of known cases). Study design The system has been validated using two independent approaches (cloning and sequencing of UL-97 gene fragments and real-time PCR) and clinical samples derived from 3 pediatric patients. Results The UL97 pyrosequencing analysis has indicated a significant increase of mutant viruses carrying the H520Q and C592G mutations. In particular, the H520Q viral mutation, known to increase GCV resistance (IC50 = 10) increased around 5 times during hospitalization. In addition, C592G (known to have IC50 = 2.9) also increased 3 times. Conclusions PSQ is a quick, cheap, high throughput and sensitive analysis method to detect GCV-associated resistance mutation useful to follow antiviral therapy in perinatal CMV-infection as well as in immune-suppressed patients. |
| Databáze: | OpenAIRE |
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