ISOLATION IDENTIFICATION AND PATTERNS OF ANTI-TUBERCULOSIS DRUG RESISTANCE FROM TERTIARY CARE HOSPITAL UDAIPUR (RAJASTHAN)
Authors: Agnihotri Sakshita , Yadav Kuldeep 
Published: August 2022

Objective: Resistance against Mycobacterium tuberculosis (MTB) is important in the sense that it has an
implication in the control of tuberculosis. The terms used to describe resistance to antituberculosis drugs
are resistance among new cases (or primary resistance) and resistance among previously treated patients. The resistance
among previously treated patients may be due to faulty treatment like prescription of inadequate treatment regimens,
interrupted availability or poor quality of drugs, or incomplete treatment adherence while subsequent transmission of these
resistant organisms to others will lead to development of disease which is resistant from the beginning called primary
resistance. Pakistan is ranked eighth in terms of global estimated burden of tuberculosis cases. Multi-Drug Resistant (MDR)
tuberculosis among new cases and MDR among previously treated patients is 3.2% and 35% respectively.
Material and methods: -
AFB smear examination and grading: - AFB smear examination was carried out by direct microscopy using the Ziehl Neelsen
(ZN) method. Sputum smear result was examined and interpreted according to the AFB grading.
AFB culture and drug susceptibility test: - Culture examinations were done on all diagnostic specimens of AFB smear
positivity. Sputum specimens from each patient were processed with sodium hydroxide (NaOH) method-Modied Petroff 's
procedure and cultured on Lowenstein-Jensen (LJ) slopes.10 All inoculated LJ drug and control media were incubated at 37ºC.
All cultures were examined 48-72 hours after inoculation to detect gross contaminants. Thereafter, cultures were examined
weekly, up to eight weeks on a specied day of the week.
Typical colonies of M. tuberculosis were rough, crumbly, waxy, non-pigmented (buff coloured) and slow-growers, i.e., only
appeared two to three weeks after inoculation. The colony was conrmed by ZN staining. Detection time for MOTT was 25 days.
M. tuberculosis positive strains were culture negative when they grew on p-nitro benzoate (PNB) containing medium. Only a few
colonies of non-tuberculosis Mycobacteria (NTM – often pigmented, with smooth morphology or PNB positive) were grown as
visible colonies on PNB containing medium.
Anti-TB drug susceptibility testing: - anti-susceptibility testing perform on pre-formed LJ media with antitubercular drugs
Tuberculosis First Line Kit (Total 7 slants) Containing ve antitubercular agent (Isoniazid, Streptomycin, Ethambutol,
Rifampicin and Pyrazinamide) 2 controls without any antimicrobial agent.
Results: out of 119 samples antitubercular testing against rst line antitubercular drugs such as Pyrazinamide were shows 12
(10.08%) sample were resistance which accounts maximum resistance among rst line antitubercular another rst line
antitubercular drugs shows resistance as follows Streptomycin (9.24%), Ethambutol (8.40%), Isoniazid (7.56%), Rifampicin
(6.72%), drugs out of 119 samples in which 107 samples were susceptible to the Pyrazinamide drug in in-vitro antitubercular
susceptibility testing. Antitubercular resistance against second line antitubercular drugs were shows as follows out of 119
samples antitubercular testing Ethionamide were shows 9 (8.18%) sample were resistance which accounts maximum
resistance among second line antitubercular another second line antitubercular drugs shows resistance as follows
Clarithromycin (6.72%), Ciprooxacin (5.88%), D- Cycloserine (5.88%), Amikacin (5.04%), Kanamycin (4.20%), P-
aminosalicylic acid ( 4.20%) and Rifabutin (3.36%) drugs out of 119 samples in which 107 samples were susceptible to the
Pyrazinamide drug in in-vitro antitubercular susceptibility testing. MDR-TB emerged in patients who were resistant to
Rifampicin and Isoniazide was 6 in number during this study.