FLUARRAY

Development and validation of a microarray based automated diagnostic system for the detection of influenza virus types and subtypes at point-of-care

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 Obiettivo del progetto (Objective)

'Influenza is an extremely contagious infection that is caused by distinct virus types and subtypes. Early diagnosis is crucial for disease treatment and control as it reduces the inappropriate use of antibiotics and provides the indication for antiviral therapy. Rapid diagnosis is also a key component of surveillance activity. This requires the ability to detect and accurately diagnose infection at or close to the source/outbreak with minimum delay, a tactic consistent with the global experience during the SARS epidemic in 2003. This experience underlines the need for specific, sensitive point-of-care testing capable of discriminating between influenza subtypes. None of the available influenza diagnostic assays combines a point-of-care format with the multiplex capability to identify a large repertoire of human and animal viruses. This project exploits the knowledge and the expertise of the partners to convert microarray assays, that have a powerful multiplex capability but are laborious, complex and expensive to perform, into a simple, robust and affordable automated point-of-care system for the diagnosis of influenza. The system will utilize three components: 1) a microarray immunoassay that distinguishes influenza A and B virus as well as A subtypes; 2) an innovative self-contained disposable lateral-flow device that allows the addition of specimens and reagents in a temporally-controlled manner; 3) a robust automated processing reading instrument of novel conception that employs a low cost, high performance optical module. This project will provide small laboratories, health offices, veterinary clinics and outposts (airports) with the diagnostic capability of major research institutions and reference centres, thus providing better care for patients and most importantly, facilitating the implementation of surveillance activities and guiding response measures that are being built to face a possible influenza pandemic caused by a highly virulent virus.'

Introduzione (Teaser)

A timely diagnosis of influenza is the key to prompt treatment. A European consortium worked on developing an innovative diagnostic tool based on microarrays.

Descrizione progetto (Article)

Over the past years, a number of influenza diagnostic tools have been developed for point-of-care applications. They are based on dipstick lateral flow technology that utilises robust, affordable and easy to perform assays. However, rapid tests have limitations, such as lack of sensitivity, an inability to distinguish among different strains of influenza viruses and little multiplex capacity.

New test formats and technologies are required for determining numerous parameters simultaneously. In this context, the EU-funded http://www.fluarray.eu/ (FLUARRAY) (Development and validation of a microarray based automated diagnostic system for the detection of influenza virus types and subtypes at point-of-care) project proposed to develop a microarray-based system for point-of-care influenza diagnosis. This technology would dramatically increase assay throughput while reducing cost.

Microarray assays are currently processed manually and require equipment not suitable for the clinical laboratory. FLUARRAY converted the sophisticated microarray technology into a simple and robust diagnostic system. This system can simultaneously process hundreds of assays on a surface area the size of a fingernail. It will be used to distinguish between haemagglutinin (HA) and neuraminidase (NA) variants from major human and animal viruses in biological specimens.

The system essentially comprises a miniature immunoassay utilising a microarray of capture antibodies directed against the two influenza virus proteins, HA and NA. The array captures recombinant tracer proteins which are then sandwiched by a secondary fluorescently-labelled antibody. In the presence of influenza virus in the specimen, the tracer proteins are displaced by the viral antigens, causing a reduction in the recorded fluorescence signal.

The consortium has produced all the molecular components of the assay including the monoclonal antibodies and assessed its detection sensitivity. Scientists have validated the performance of the system but the desired sensitivity is yet to be achieved.

Considerable efforts were dedicated towards the production of a prototype automated microarray reading instrument that can add samples and reagents to the assay device. The instrument contains an optical module to read microarray immunoassays with a sensitivity and dynamic range similar to that of the most sophisticated microarray scanners.

FLUARRAYs portable point-of-care diagnostic system will perform hundreds of distinct parallel immunoassays offering high discriminatory capacity in the diagnosis of influenza.