Key result
Pre-amplification of clinical samples using human airway epithelium cultures followed by nested PCR increased the detection rate of respiratory viruses from 76.5% to 100%.
Why the study?
Does pre-amplification in human airway epithelium cultures improve the detection of respiratory viruses in clinical samples from adult patients with asthma exacerbations?
Observational (n=34)
No
Does pre-amplification in human airway epithelium cultures improve the detection of respiratory viruses in clinical samples from adult patients with asthma exacerbations?
Absolute Event Rate: 100% vs 76.47%
Pre-amplification of clinical samples in human airway epithelium cultures combined with broad-spectrum nested PCR significantly improves the diagnostic yield for respiratory viruses, identifying pathogens in previously undiagnosed cases.
May aid diagnosis of respiratory infections; leaves open clinical validation before routine use.
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.
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Pyrć et al. (2012) conducted an observational in Respiratory tract infections in asthma exacerbations (n=34). Human airway epithelium (HAE) cultures followed by nested PCR vs. Nested PCR alone was evaluated on Detection of respiratory viruses. Pre-amplification of clinical samples using human airway epithelium cultures followed by nested PCR increased the detection rate of respiratory viruses from 76.5% to 100%.
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