Development of a RT Real-Time PCR for the Detection and Quantification of Human Rhinoviruses

Gambarino, Stefano; Costa, Cristina; Elia, Mariateresa; Sidoti, Francesca; Mantovani, Samantha; Gruosso, Valentina; Bergallo, Massimiliano; Cavallo, Rossana

Human Rhinoviruses (HRV) are the most common viral agents, being responsible for upper as well as lower respiratory tract infections. Evidence demonstrating that HRV disease is not exclusively limited to the upper airways and may cause lower respiratory complications, together with the frequency of HRV infections and the increasing number of immunocompromised patients underline the need for including HRV in virological diagnostics of acute lower respiratory tract illness. This article describes the development and optimization of a reverse transcription (RT) real-time PCR assay for quantification of HRV RNA in clinical samples. Efficiency, sensitivity, specificity, inter- and intra-assay variability, and dynamic range have been determined. Subsequently, the assay has been validated on bronchoalveolar lavage (BAL) specimens obtained from immunocompetent and immunocompromised patients.