DEMO-NOPERSIST

"Demonstration of NOPERSIST results leading to novel, validated diagnostic tests for active human and bovine tuberculosis"

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 Obiettivo del progetto (Objective)

'Tuberculosis (TB) in humans and bovine TB in farm animals are global health problems of immense social and economic importance.

Human TB: Mycobacterium tuberculosis (M.tb) is a slowly replicating bacillus that resides intracellularly within phagosomes of macrophages and commonly causes latent infections of the lung and in about 5% of the infected individuals it leads to active disease. Co-infection with M. tuberculosis is estimated in about one-third of HIV-1 infected subjects. Indeed, the risk of developing M. tuberculosis as an opportunistic infection is increased up to 200-fold in HIV-1 subjects. Drug resistance to HIV-treatment and appearance of multiple-drug resistance (MDR) and off late of Extra-Drug Resistance (XDR) strains of M. tuberculosis, the causative agent of human TB is steadily leading to a hopeless situation as far as the therapy is concerned.Bovine TB: Bovine tuberculosis is caused by the bacterium Mycobacterium bovis, which is closely related to Mycobacterium tuberculosis, the cause of human tuberculosis. M. bovis can be transmitted to humans through the ingestion of unpasteurised milk and milk products as well as aerosols, from infected cows. The introduction of pasteurisation eliminated transmission through contaminated milk and greatly reduced the human health problem1. The principal hosts for bovine tuberculosis are cattle and buffalo, however many other domestic and wild animals can become infected e.g. goats, cervids, pigs, wild boars, dogs, cats, camels, badgers, primates, hares, amongst others. Bovine tuberculosis is found worldwide. All developed countries currently have a TB eradication program in place for many years. These programs have been largely successful; however, incidences are increasing in many countries (e.g. UK, Ireland, France, Austria, Germany). The specific objectives achievable within DEMO-NOPERSIST project are

1. Development and evaluation of prototype test kits for active human TB (LIONEX, SME). This test shall be the World’s first blood test for discriminating latent from active TB in humans.

2. Development and evaluation of prototype test kits for active Bovine TB (PRIONICS, SME)

3. To develop marketable, improved diagnostic products for human and bovine TB within a period of 2-3 years .

Thus, both human and bovine active TB tests are directly linked to the NOPERSIST project and shall be an excellent demonstration of a Res4SME project NOPERSIST.'

Introduzione (Teaser)

Human and bovine tuberculosis (TB) are global health problems of immense social and economic importance. Urgently needed diagnostic methods capable of differentiating between active and latent infections are under development.

Descrizione progetto (Article)

TB, affecting one third of the human population, is caused by Mycobacterium tuberculosis (MTb). TB is a leading killer of people infected with HIV, and the appearance of multi-drug-resistant strains of MTb further complicates the problem. Currently, no efficient and cost-effective tests are available for early diagnosis of TB, and there are no tests that can differentiate millions of infected people from active TB patients.

Closely related bacteria M. bovis causes bovine TB, found worldwide. The principal hosts for bovine TB are cattle and buffalo, however many other domestic and wild animals can become infected.

The DEMO-NOPERSIST (Demonstration of NOPERSIST results leading to novel, validated diagnostic tests for active human and bovine tuberculosis) project aims to produce marketable diagnostic products for TB in people and cattle, and also to differentiate between latent and active infections in people. This two-year project, to be completed in September 2015, puts the results of the previous EU project NOPERSIST into practice.

Stages of disease can be assessed through interferon gamma (IFN-gamma) and interleukin 2 (IL-2), secreted by the T helper cells of the immune system. To develop diagnostics, the partners already selected and are producing recombinant antigens from MTb. They are developing the monoclonal antibodies, anti-IL-2 and anti-IFN-gamma, which will be used for kit production.

Human IFN-gamma and IL-2 tests are developed in several formats. The ELISA test contains 96-well microtiter plate, coated by IL-2- and/or IFN-gamma-specific antibodies, ready-to-use detection antibody, conjugate and the necessary buffers. The cytokine release assay (human ELISPOT test) contains ready-to-use stimulating antigens, coating and detection antibodies, streptavidin-horseradish peroxidase conjugate and substrate solution. A bovine TB diagnostic is being developed in a lateral flow format.

The ability to differentiate between latent and active TB is of extreme importance, considering a large percentage of the infected population remains otherwise healthy. The developed diagnostics will have an immense positive impact on human health and prevention of TB.