Microbiology Department, Faculty of Biochemistry Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
PLoS One. 2012;7(3):e32582. doi: 10.1371/journal.pone.0032582. Epub 2012 Mar 5.
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.
快速准确地检测和鉴定引起呼吸道感染的病毒对于患者的治疗和疾病的控制非常重要。尽管有几种检测方法可用，但仍有约 30%的呼吸道疾病患者无法确定病因。因此，本研究的目的是开发一种诊断试剂盒，用于检测灵敏度低至 1-10 拷贝/反应的呼吸道病毒。使用训练临床样本集评估该检测方法显示，约 76%的病例中可鉴定出病毒核酸。为了提高检测性能并有助于鉴定新的病毒种类或新兴株系，使用完全分化的人呼吸道上皮细胞培养物对传染性病毒进行预扩增。这一额外步骤使得所有测试样本均能检测到病原体。基于这些结果可以假设，一些临床样本中缺乏病因，这既包括之前报道的，也包括本研究中观察到的，其原因不仅可能是未知病毒种类的存在，还可能是由于所用检测方法的不完善。
