Use of Sensitive, Broad-Spectrum Molecular Assays and Human Airway Epithelium Cultures for Detection of Respiratory Pathogens
| Autor: | Jan Potempa, Andrzej Szczeklik, Slawomir Zeglen, Krzysztof Wojcik, Marian Zembala, Wojciech Karolak, Krzysztof Pyrc, Marek Ochman, Grazyna Bochenek, Marek Sanak, Katarzyna Gawron, Magdalena Hubalewska-Mazgaj, Karol Stożek, Jacek Wojarski |
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| Rok vydání: | 2012 |
| Předmět: |
Viral Diseases
Pulmonology Respiratory System Cell Culture Techniques lcsh:Medicine Biology medicine.disease_cause Microbiology Polymerase Chain Reaction Epithelium law.invention Diagnostic Medicine law Virology Influenza A virus medicine Humans Respiratory system lcsh:Science Polymerase chain reaction DNA Primers Multidisciplinary Respiratory tract infections lcsh:R Infectious Diseases medicine.anatomical_structure Cell culture Culture Media Conditioned Respiratory Infections Viruses Immunology Nucleic acid Medicine lcsh:Q Research Article Respiratory tract |
| Zdroj: | PLoS ONE, Vol 7, Iss 3, p e32582 (2012) PLoS ONE |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0032582 |
| Popis: | Rapid and accurate detection and identification of viruses causing respiratory tract infections is important for patient care and disease control. Despite the fact that several assays are available, identification of an etiological agent is not possible in ~30% of patients suffering from respiratory tract diseases. Therefore, the aim of the current study was to develop a diagnostic set for the detection of respiratory viruses with sensitivity as low as 1-10 copies per reaction. Evaluation of the assay using a training clinical sample set showed that viral nucleic acids were identified in ~76% of cases. To improve assay performance and facilitate the identification of novel species or emerging strains, cultures of fully differentiated human airway epithelium were used to pre-amplify infectious viruses. This additional step resulted in the detection of pathogens in all samples tested. Based on these results it can be hypothesized that the lack of an etiological agent in some clinical samples, both reported previously and observed in the present study, may result not only from the presence of unknown viral species, but also from imperfections in the detection methods used. |
| Databáze: | OpenAIRE |
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