Publishable summary: 

Tuberculosis (TB) is a devastating disease affecting millions of people around the globe. The increasing appearance of multi-drug-resistant (MDR) strains worsens the threat to human kind with unprecedented risks. Rapid and cost-effective methods for early diagnostics and detection of MDR are required. Pyrazinamide (PZA) is the only drug that effectively kills dormant tuberculosis. In average two-thirds of the world population is infected with latent TB. Resistance to PZA is increasing globally and no other drug has yet been identified to replace PZA for resistant strains. Therefore, it is necessary to better understand the mechanisms of action and resistance of PZA to put the basis towards the design of alternative drugs. It is also important to early and accurately detect PZA-resistance in order to improve TB care and control. Here, we form a consortium with expertise in biochemistry, biophysics, bioinformatics, genetics and clinical research, aiming to develop novel methods for early diagnosis and real-time monitoring of TB infections and PZA resistance markers.

Two main streams of collaborative research will flow along the EU- LAC axis:

  1. Detection of Mycobacterium tuberculosis by single-cell Raman spectroscopy on Lab-on-aChipdevices with enhanced materials for cell isolation from complex biological samples, in particular from direct sputum.
  2. Detection of PZA-resistant M. tuberculosis markers by exploiting the potential and versatility of molecular sensing combined with novel nanomaterials on inexpensive sensors.

The MYCO-NET2 consortium will deploy state of art technologies to allow Lab-on-a-Chip and nanotechnology for TB detection and PZA resistance determination to be used in developing countries as a point-of-care diagnostics in a sustainable manner. So far, in tihis first year we obtained approximately 500 sputum samples from TB patients and controls, and cultured them in presence of PZA. Antibodies and aptamers against POA and specific proteins of MTB have been produced. Micro-Raman spectra for MTB and POA have been obtained and confirmed its validity for detecting these specimens. Advances in the nanostructured filters are being done with promising results.