The detection of influenza A and B viruses in clinical specimens using a quartz crystal microbalance

Abstract

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.

Real-time recognition of Mycobacterium tuberculosis and lipoarabinomannan using the quartz crystal microbalance

“…While ELISA is normally considered a gold standard, the QCM has been found to be more sensitive in some instances. [12] ELISA benefits from natural enzymatic signal amplification, while QCM is faster with real-time results relying on the densities of the adsorbed particles for signal amplification. The ELISA results utilized whole cells which were attached to the well plate first.…”

“…[11] An immunosensor based on the QCM meets these requirements and has been used as a platform for diagnostic immunoassays. [12–14] These assays can be adapted for use with antibodies to multi-drug resistant and extremely-drug resistant tuberculosis, reducing the time it takes to develop diagnostics for new strains of tuberculosis. Since current analytical techniques have not sufficiently demonstrated, practicality for use at the POC, the use of the QCM to detect Mtb bacilli and antigens will be assessed as a competing technology.…”

Detection of viruses using piezoelectric cantilevers

“…Самыми распространенными из них яв ляются микровесы на основе к варцевого криста лла [7,8,9], а так же дат чик на основе эффекта поверхностных акустических волн в пьезокри-сталле [10]. В новых разработках так же пред-лагается использовать емкостные микроме-ханические ультразвуковые преобразователи [11] и мембранные пьезоакустические резона-торы [12].…”

“…N u m e r o u s p u b l i c a t i o n s describe the use of various resonant sensors for virus detection. The most common sensors are quartz crystal based microbalances [7,8,9] and sensors based on the effect of surface acoustic waves in a piezoelectric crystal [10]. In new developments, it is also proposed to use capacit ive m ic romecha n ica l u lt rasonic transducers [11] and membrane piezo-acoustic resonators [12].…”

QCM immunosensor detection of Escherichia coli O157:H7 based on beacon immunomagnetic nanoparticles and catalytic growth of colloidal gold

“…Thereafter, it was used for the detection of other microor-0956-5663/$ -see front matter © 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.bios.2010.12.035 ganisms, including Bacillus anthracis, Schistosoma japonicum, and the influenza virus (Hao et al, 2009;Wang et al, 2006;Peduru et al, 2009). However, these QCM biosensors have a high detection limit of 10 5 -10 7 CFU/ml for bacteria.…”