Open AccessUse of Sensitive, Broad-Spectrum Molecular Assays and Human Airway Epithelium Cultures for Detection of Respiratory Pathogens
Author(s) -
Krzysztof Pyrć,
Karol Stożek,
Krzysztof Wójcik,
Katarzyna Gawron,
Sławomir Żegleń,
Wojtek Karolak,
Jacek Wojarski,
Marek Ochman,
Magdalena Hubalewska-Mazgaj,
Grażyna Bochenek,
Marek Sanak,
Marian Zembala,
Andrzej Szczeklik,
Jan Potempa
Publication year - 2012
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0032582
Subject(s) - respiratory tract , respiratory tract infections , biology , respiratory epithelium , respiratory system , virology , nucleic acid , etiology , microbiology and biotechnology , immunology , medicine , pathology , genetics , anatomy
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.