Project: Development of an innovative kit for the early screening of waterborne pathogens

Viruses flourish in water environments. _x000D_Over 140 virus species which impact human health are present in sewage-contaminated waters (World Health Organization- WHO 2010). _x000D_These waterborne pathogens include human enteric viruses that cause gastroenteritis (e.g. norovirus, sapovirus, adenovirus, astrovirus, rotavirus), hepatitis (A and E) and other illnesses such as fever or respiratory infections. _x000D__x000D_Among human enteric viruses, about 1.4 million cases of hepatitis A occur annually worldwide (WHO, 2008). It may take months for patients to recover from the debilitating symptoms and to be well enough to resume normal daily life._x000D_Similarly, noroviruses are estimated to cause 23 million cases of acute gastroenteritis in the United States each year and at least 50% of foodborne illness outbreaks (ECDC, 2009)._x000D__x000D_Human enteric viruses epidemics can also be explosive in growth: the largest recorded waterborne outbreak of this kind involved 300 000 cases of Hepatitis A in Shanghai in 1988. In Europe, the average number of outbreaks of waterborne diseases is estimated to be between 35 and 50 per year (WHO-ENHIS’s LBE, 2009). _x000D__x000D_Human enteric viruses enter the water environment through sewage-contaminated water. Water treatment practices may fail to ensure the full removal of viral pathogens if there are deficiencies in treatment processes or in water distribution. Then enteric viruses may be transmitted to humans through a number of different routes, such as via shellfish grown in contaminated waters, food crops grown in land irrigated or fertilized with sewage water and sludge, sewage-polluted recreational waters or contaminated drinking water._x000D__x000D_Irrespective of the sanitation issues, the detection of human enteric viruses in the water environment is a major issue to COtain the health of the population. _x000D__x000D_Currently, the detection of viruses in water is COly performed via either quantitative polymerase chain reaction (PCR) methods (in vitro DNA amplification techniques that allow multiplication of specific DNA sequences) or cell culture. Yet both methods have drawbacks when faced with the analysis of water samples of large volume:_x000D_- Highly-skilled staff are required,_x000D_- The turnaround time to obtaining a test result can be long (2 days for PCR and 1 month for cell culture),_x000D_- Cost are high (150-300€ per analysis and per virus for PCR and 230-400€ for cell culture detection of enteric viruses)._x000D__x000D_In the face of these limitations there is a need for screening solutions that would allow water supply managers to detect a viral threat early and on site, through routine use of diagnostic kits._x000D_The use of high-throughput screening kit on site would be a breakthrough in the water industry._x000D__x000D_This is the precise goal of the ENVIRUS project aiming to develop a new assay kit that will enable rapid, flexible quantification of viruses in water samples. It will use nanomaterials with remarkable virus recognition properties (virus imprinted particles,VIP) for the pre-concentration of viruses in VIP-packed cartridges and their subsequent detection in VIP-coated microplates. The use of international standard 96-well microplates as the base for the ENVIRUS kit provides the assay with intrinsic flexibility. It enables the design of different kits to meet varying analysis demands rapidly, ranging from multiplex screening applications (e.g. allowing 4 different viruses to be analysed in 6 triplicate samples in only a few hours) to high-throughput applications (allowing analysis of 96 samples for one single virus)._x000D__x000D_And the target price is attractive at between 10 and 20€ per sample._x000D__x000D_The ENVIRUS kit will allow industry, farmers and other public sanitary authorities to self-monitor their water environments. This degree of active viral risk management holds the potential to have a substantial positive impact on public health, with cost benefits in terms of expenditure on testing, monitoring and disinfection._x000D__x000D_There is no solution of this kind on the market today._x000D__x000D_The ENVIRUS project is initiated by:_x000D_- Two-spin-out companies_x000D_. ENVOLURE (ENV): A French company specializing in the characterisation of residual waters through a new microplate-based detection technology, _x000D_. INOFEA (INOF): A Swiss company producing innovative nanomaterials for the specific binding of viruses._x000D_- The University of Applied Sciences Northwestern Switzerland (FHNW): Expert in molecular nano-biotechnology and owner of the patent underlying the project,_x000D_- VEOLIA: A French company recognised to be the world’s biggest player in the management of water services. The company’s research teams have are strong expertise in the detection of pathogens in water._x000D__x000D_The 36-month work programme (maximum duration) will target the screening of enteric viruses in drinking and industrial waters and then in sewage waters._x000D_

Acronym	ENVIRUS (Reference Number: 6894)
Duration	01/03/2012 - 27/02/2015
Project Topic	Development of a new kit for the early screening of waterborne pathogens (firstly human enteric viruses carrying gastroenteritis and hepatitis A and E). The kit will show high sensitivity and faster laboratory turnaround time than current detection methods, at lower cost
Project Results (after finalisation)	The ENVIRUS project was structured as a puzzle, with five pieces needing to be developed and assembled to come up with a marketable product (virus detection kit for waters): (1) design of nanoparticles with virus recognition properties, (2) design of a detection system, (3) binding of the nanoparticles in the 96-well microplate to generate the assay kit, (4) adapt the assay to waters, and (5) understand the market to properly address it with a well-adapted product._x000D__x000D_The CO results of our project are a proof of technical feasibility and the absence of an appropriate market for the tentative product. On the technical side, nanoparticles with norovirus recognition properties were properly designed and shown to be efficient (1), a highly innovative detection system was designed and coupled directly to the nanoparticles (2), binding of the nanoparticles to the microplate was achieved (3), and the detection of norovirus was demonstrated in water. Therefore the various individual technical steps were achieved. However, on the market side (5), we came to understand that the initially targeted market consisting of on-site water-treatment and water-production operators are not responsive to the tentative product. The pre-concentration of viruses required before analysis introduces a risk that the operators are not willing to take. The surrogate market could be the environmental analysis laboratories, but in this case the tentative product is in direct competition with RT-PCR commonly used in those labs.
Network	Eurostars
Call	Eurostars Cut-Off 7

Project partner

Number	Name	Role	Country
4	ENVOLURE	Coordinator	France
4	Fachhochschule Nordwestschweiz - School of Life Sciences	Partner	Switzerland
4	INOFEA	Partner	Switzerland
4	VEOLIA ENVIRONNEMENT RECHERCHE ET INNOVATION	Partner	France