Immuno-interferometric sensor for the detection of influenza A nucleoprotein

Abstract

Influenza A virus, both seasonal and pandemic, has the potential to cause rampant devastating disease around the world. The most relied upon methods of viral detection require days, skilled workers, and laboratory settings to complete properly. Here, we report two methods for the detection of the nucleoprotein from inactivated influenza A (IFA-NP), a patented polymer-protein antibody orientation immuno-method, termed ALYNGSA, and a newly fabricated optical label-free Fabry-Perot interferometric immunosensor. The ALYGNSA assay for IFA-NP had a level of detection below 5 microg/mL of inactivated virus sample. The label-free detection through Fabry-Perot interferometry with the ALYGNSA orientation yielded an improved sensitivity to 1 microg/mL over the fluorescence sandwich assay alone. Characterization of the detection surface by fluorescence microscopy and non-contact AFM corroborated interferometry results. The resulting label-free detection method has the prospective for adaptation into a portable multi-chip sensor capable of real-time in situ detection of influenza A virus.

An Immunosensor Based on Antibody Binding Fragments Attached to Gold Nanoparticles for the Detection of Peptides Derived from Avian Influenza Hemagglutinin H5

“…Immunosensors have found widespread applications in food industry, environmental and pollutants, biotechnology, pharmaceutical chemistry and clinical diagnostics [ 22 – 27 ]. Several types of immunosensors, such as surface plasmon resonance (SPR) [ 3 , 28 , 29 ] and quartz crystal microbalance (QCM) [ 30 – 34 ], optical interferometric [ 35 , 36 ], label-free microcantilever [ 37 ] and fluorescent [ 38 ] have been researched as alternatives to the conventional influenza virus detection methods. These biosensors have shown analytical potential, but they lack subtype specificity.…”

Extrinsic surface-enhanced Raman scattering detection of influenza A virus enhanced by two-dimensional gold@silver core–shell nanoparticle arrays

“…Finally, comparison of the LOD with those of other techniques previously reported for detecting the same analyte. The LOD of our SERS immunoassay system (8 ng mL À1 ) is much lower than those of uorescence-based LFIA (250 ng mL À1 ), 26 an immune-interferometric sensor (1 mg mL À1 ), 27 and comparable to that of electrochemical immunoassay (e-ELISA) (10 ng mL À1 ). 28 3.3.3 Selectivity.…”

Simple and inexpensive immunoassay-based diagnostic tests

“…The device includes also moving stages, which makes detection of multiple chips possible. The feasibility of the sensor has been demonstrated with influenza A nucleoprotein assay reaching detection limit of 1 μg/ml [74].…”

“…While most of the assay chips are very small in size and often based on microfluidics requiring thus only minute volumes of reagents and sample, the instrumentation needed for quantification is still rather large [29,74], or portable desktop devices [26,70,71] and very few handheld systems exist [15,34]. Also, a computer is often needed [74,80], which raises the costs of the device and increases the size of the instrumentation needed. Therefore, miniaturization of instrumentation towards truly handheld devices or development of totally instrument-free quantitative assays is the key issue.…”