Abstract

Objective: Our objective of the work was to identify the type of Mycobacterium at species level with an aim to diagnose and treat the resistant strains by using MDR strains in infecting in patients to identify the infecting Mycobacterium and to find resistance at the level of genes tested using molecular biology of line probe assay methods.

Design: We tested various samples for Mycobacterium by using microbiology and molecular biology method of TB-TMA from clinical suspects. With use of Amplification of gene and line arrays of bacteria we determined the susceptibility character of drugs of infecting Mycobacteria in the patients.

Results: We were able to identify Mycobacterium tuberculosis by measuring the activity at Promoter level of Mycobacteria and determined their first and send line of drug sensitivity using molecular biological tools of drug susceptibility of MDR and XDR sure methods developed by us. We could detect mutations in repo B gene imparting drug resistance to Rifampicin or Kat G gene and In A gene to Isoniazid from patients’ samples infected with Mycobacterial species or resistance to fluoroquinolone, by detecting mutations in gyrA gene and rrs gene that imparts resistance of bacteria to injectable antibiotics. We were able to also Identify Mycobacterium Other Then Tuberculosis (MOTT)

Material and methods

Samples: We obtained samples of clinical TB suspects from various multispecialized hospitals in our Norquest laboratory at Central lab, Indore (M.P). These patients were recorded in the hospitals and tested using medical, for treatment of patients. The clinical suspects were tested by medical specialists to have pulmonary or extra pulmonary TB, tested by medical methods like chest radiography, C.T scan, pleural tap and CBC tests and dispatched. The samples were transported immediately to our laboratory under cold temperature conditions. Both the pulmonary as well as extra pulmonary samples were analyzed in our laboratory for diagnosis of tuberculosis. For Drug. Sensitivity test three consecutive early morning sputum samples were collected by laboratory technologist, according to protocol in our NABL accredited, Central lab-Norquest, Indore. The samples were processed on the same day or were kept at +4ᵒC till processed. The Medical diagnosis of suspects was done by medical specialists. And M. tuberculosis was detected using TB-TMA method to identify the infecting bacteria as described earlier.

Conclusion: We found great variations in the genes of Mycobacterium infecting patients tested in our lab. We were able to find drug sensitivity or resistance of each bacteria infecting, which proved out to be very useful for Clinicians for prescription of the drug regimen based on the infecting bacteria in individual patient according to type of species the patient is infected with Multidrug-resistant tuberculosis (MDR-TB) is tuberculosis disease caused by organisms, which show different level of resistance to Isoniazid or Rifampicin, which may not show resistance to other anti-TB drugs. With increase in MDR TB around the world, among new cases, as well as in the previously treated ones as reported by WHO (2010) [1] & [2].