The detection of influenza A and B viruses in clinical specimens using a quartz crystal microbalance.
Current methods for the accurate diagnosis of influenza based on culture of the virus or PCR are highly sensitive and specific but require specialised laboratory facilities and highly trained personnel and, in the case of viral culture, can take up to 14 days to obtain a definitive result. In this study, a quartz crystal microbalance-based immunosensor (QCM) has been developed and its potential evaluated for the rapid and sensitive detection of both influenza A and B viruses in laboratory-cultured preparations and clinical samples. The effective limit for detection by QCM for stock preparations of both A/PR/8/34 and B/Lee/40 viruses was 1 x 10(4) pfu/mL, associated with observed frequency shifts of 30 (+/-5) and 37 (+/-6.5) Hz, respectively. Conjugation of 13 nm gold nanoparticles to the detecting antibody improved the mass sensitivity of the immunosensor, resulting in a 10-fold increase in sensitivity and a detection limit of 1 x 10(3) pfu/mL for both preparations, with resulting frequency shifts of 102 (+/-11) and 115 (+/-5) Hz, respectively. Detection of virus in nasal washes with this technique was achieved by overnight passage in MDCK cultures prior to analysis. A comparison of results obtained from 67 clinical samples using existing RT-PCR, shell vial, cell culture and ELISA methods showed that QCM techniques were comparable in sensitivity and specificity to cell culture methods.
Peduru Hewa
Thamara M
TM
School of Applied Sciences, Royal Melbourne Institute of Technology, Melbourne 3001, Australia.
Tannock
Gregory A
GA
Mainwaring
David E
DE
Harrison
Sally
S
Fecondo
John V
JV
D000818:Animals; D000914:Antibodies, Viral; D015374:Biosensing Techniques / Q000295:instrumentation*; D002460:Cell Line; D003460:Crystallization; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D006046:Gold / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000502:physiology; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000502:physiology; D009981:Influenza B virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000502:physiology; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D055617:Micro-Electrical-Mechanical Systems; D011791:Quartz; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D014776:Virus Cultivation
D000914:Antibodies, Viral; D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic; D011791:Quartz; D006046:Gold