A touchdown nucleic acid amplification protocol as an alternative to culture backup for immunofluorescence in the routine diagnosis of acute viral respiratory tract infections

摘要: Immunofluorescence and virus culture are the main methods used to diagnose acute respiratory infections. Diagnosing these infections using nucleic acid amplification presents technical challenges, one of which is facilitating different optimal annealing temperatures needed for each virus. To overcome this problem we developed a diagnostic molecular strip combined generic nested touchdown protocol with in-house primer master-mixes that could recognise 12 common viruses. Over an 18 month period total 222 specimens were tested by both immunofluorescence strip. The came from 103 males (median age 3.5 y), 80 females 9 y) 5 quality assurance scheme specimens. Viruses recovered number specimen types including broncho-alveolar lavage, nasopharyngeal secretions, sputa, post-mortem lung tissue throat nasal swabs. Viral detection IF was poor in sputa A 99 viruses detected study 79 patients 4 control specimens: 31 consistently out-performed no loss specificity. pre-dispensed suitable replacing diagnosis negative immunofluorescence. Results available after average 4–12 hours while results within 24 hours, considerably faster than viral culture. proved be convenient reliable method testing multiple routine setting.