Discrimination of infectious bacteriophage T4 virus by propidium monoazide real-time PCR
| Autor: | Bárbara Adrados, Jordi Morató, Francesc Codony, Nancy J Pino Rodríguez, Mariana Fittipaldi, Gustavo A. Peñuela |
|---|---|
| Přispěvatelé: | Universitat Politècnica de Catalunya. Departament d'Òptica i Optometria, Universitat Politècnica de Catalunya. TMAS (T+) - Toxicologia i Microbiologia Ambiental i Sanitària |
| Rok vydání: | 2010 |
| Předmět: |
Azides
Hot Temperature Ciències de la salut::Medicina [Àrees temàtiques de la UPC] Polymerase Chain Reaction Sensitivity and Specificity Virus Microbiology law.invention Specimen Handling Bacteriophage chemistry.chemical_compound law Propidium monoazide Virology Bacteriophage T4 Humans Polymerase chain reaction Bacteriologia Microbial Viability biology Bacteriology biology.organism_classification Intercalating Agents Reverse transcription polymerase chain reaction Real-time polymerase chain reaction Capsid chemistry Viruses DNA Propidium |
| Zdroj: | Recercat. Dipósit de la Recerca de Catalunya instname UPCommons. Portal del coneixement obert de la UPC Universitat Politècnica de Catalunya (UPC) |
| ISSN: | 1879-0984 |
| Popis: | The advent of quantitative PCR has improved the detection of human viral pathogens in the environment. However, a serious limitation of this method may arise from the inability to discriminate between viruses that are infectious and viruses that have been inactivated and do not represent a human health hazard. To assess whether propidium monoazide (PMA) pre-treatment is a good approach to inhibiting DNA amplification from non-infectious viruses, bacteriophage T4 survival was measured using cell culture titration and real-time PCR with and without PMA pre-treatment. Heat (85 degrees C) and proteolysis methods were carried out. After these inactivation treatments, the results indicated that the PMA pre-treatment approach is not appropriate for differentiating infectious viruses. However, when a heat treatment at 110 degrees C was undertaken, PMA pre-treatment did allow differentiation of non-infectious from infectious viruses. In this case, effective binding of PMA to bacteriophage T4 DNA could be taken to indicate capsid damage. Therefore, PMA pre-treatment may be appropriate for assessing effective disinfection treatments and for a more reliable understanding of the factors that contribute to viral inactivation through capsid damage monitoring. The PMA-PCR approach could be useful as a rapid and inexpensive analytical tool for screening and evaluation of the efficacy of disinfectants. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect