Fig. 2

CRISPR-Cas based methods for the diagnosis of tuberculosis. General workflow for the detection of target DNA sequences is represented. After DNA extraction (1), sample is subjected to a nucleotide amplification step through an isothermal method, which has lower requirements of equipment compared to traditional PCR (2). Target DNA is recognized by the crRNA coding the complementary sequence which forms a crRNP with Cas12 or Cas13 molecules, leading to the activation of their trans cleavage domain (3). Synthetic reporter oligonucleotides are released from the quencher molecule after cleavage in trans by the crRNP and the fluorescent signal is read (4). RPA recombinase polymerase assay, LAMP loop-mediated isothermal amplification