Supplementing standard cell culture and serology with seminested RT-PCR increased the overall rate of respiratory pathogen identification in adult asthmatics from 28% to 57%.
Observational (n=159)
Does seminested RT-PCR improve the detection of rhinoviruses compared to cell culture in adult asthmatics with respiratory infections?
Seminested RT-PCR significantly improves the detection of rhinoviruses in clinical specimens compared to traditional cell culture methods.
Absolute Event Rate: 57% vs 28%
A seminested RT-PCR (nRT-PCR) was used to detect picornavirus (PV) RNA in cell cultures inoculated with rhinoviruses (HRVs) and enteroviruses (EVs). PCR tests in which a primary "touchdown" PCR was followed by secondary reactions using PV or HRV specific primers were able to differentiate HRVs of 48 serotypes from EVs. PVnRT-PCR and HRVnRT-PCR were then used to test nasal and throat swabs from adult subjects with naturally acquired respiratory virus infections. The swabs were also analysed for respiratory viruses by cell culture techniques and the rates of PV identification by the two methods were compared. PVnRT-PCR was found to be at least five times more sensitive than cell culture for the detection of PVs in these clinical specimens. Paired acute and convalescent serum samples were tested for complement fixing antibodies to adenovirus, influenza A and B, respiratory syncytial virus, parainfluenza viruses 1, 2, and 3, Myco plasma pneumoniae, and Chlamydia psittaci. An enzyme-linked immunosorbent assay (ELISA) was used to detect rises in antibody level to coronavirus types 229E and OC43. The overall rate of pathogen identification in 159 swabs from adult asthmatics increased from 28% when only cell culture and serology were used to 57% when these methods were supplemented by PVnRT-PCR.
Ireland et al. (Tue,) conducted a observational in Respiratory virus infections in adult asthmatics (n=159). Seminested RT-PCR (PVnRT-PCR) vs. Cell culture and serology was evaluated on Overall rate of pathogen identification. Supplementing standard cell culture and serology with seminested RT-PCR increased the overall rate of respiratory pathogen identification in adult asthmatics from 28% to 57%.