title: Comparison of two commercial molecular assays for simultaneous detection of respiratory viruses in clinical samples using two automatic electrophoresis detection systems.
abstract: Two molecular assays were compared with real-time RT-PCR and viral culture for simultaneous detection of common viruses from respiratory samples: a multiplex ligation-dependant probe amplification (MLPA) and a dual priming oligonucleotide system (DPO). In addition, the positive detections of MLPA and DPO were identified using two different automatic electrophoresis systems. A panel of 168 culture-positive and negative samples was tested by the molecular assays for the presence of influenza A and B virus, respiratory syncytial virus, human metapneumovirus, rhinovirus, coronaviruses, parainfluenza viruses and adenovirus. One hundred and twenty-nine (77%) samples were positive as detected by at least one method. Sixty-nine (41%) samples were positive by cell culture (excluding human metapneumovirus and coronaviruses), 116 (69%) by RT-PCR, 127 (76%) by MLPA and 100 (60%) by DPO. The MLPA yielded results in one attempt for all samples included while 12 (7.2%) samples had to be repeated by the DPO assay due to inconclusive results. The MLPA assay performed well in combination with either electrophoresis system, while the performance of the DPO assay was influenced by the electrophoresis systems. Both molecular assays are comparable with real-time RT-PCR, more sensitive than viral culture and can detect dual infections easily. Results can be obtained within 1 day.
authors.affiliation: Laboratory of Medical Microbiology and Infectious Diseases, Isala Clinics, Stilobadstraat 3, 8021 AB Zwolle, The Netherlands. e.s.bruijnesteijn@isala.nl
authors.forename: L E S
authors.initials: LE
authors.lastname: Bruijnesteijn van Coppenraet
authors.forename: C M A
authors.initials: CM
authors.lastname: Swanink
authors.forename: A A
authors.initials: AA
authors.lastname: van Zwet
authors.forename: R H T
authors.initials: RH
authors.lastname: Nijhuis
authors.forename: J
authors.initials: J
authors.lastname: Schirm
authors.forename: J A
authors.initials: JA
authors.lastname: Wallinga
authors.forename: G J H M
authors.initials: GJ
authors.lastname: Ruijs
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001331:Automation / Q000379:methods*; D002648:Child; D002675:Child, Preschool; D004586:Electrophoresis / Q000379:methods*; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*; D014776:Virus Cultivation; D014777:Virus Diseases / Q000175:diagnosis* / Q000821:virology; D014780:Viruses / Q000302:isolation & purification*; D055815:Young Adult