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An acute respiratory syndrome Corona Virus 2 has affected humanity throughout the world. Scientifically, Corona Virus 2 is known as SARS-COV-2 which is abbreviated as COVID-2019. China was the first victim of this outbreaks in December 2019 [1] which was later recognized as pandemic on March 11, 2020 by World Health Organization (WHO) [2,3]. At the time of this writing, about 8.75 million individuals of 188 countries [4] have been effected by COVID resulting in 463000 deaths primarily, corona virus communicates from one body to another body through close contacts via droplets produced by sneezing, coughing or taking by infected badly within a buffer zone of 3 to 6 feet [5,6,7]. These droplets fall onto a surface and can survive up to 72 hours [8]. Various studies have proved that droplets may travel up to 37 feet by an uncovered cough [9,10,11]. Corona virus is not an airborne, however it may transport through respiratory droplets during talking and breathing [12].

The pncA gene encodes pyrazinamidase enzyme which converts drug pyrazinamide to active form pyrazinoic acid, but mutations in this gene can prevent enzyme activity which leads to pyrazinamide resistance. The cross‐sectional study was carried out during 2016–2017 for 12 months. The purpose of the study was to detect mutation at codon 12 and codon 85 in the pncA gene in local multidrug‐resistant tuberculosis (MDR‐TB) patients by developing a simple molecular test so that disease could be detected timely in the local population. DNA extracted from sputum‐cultured samples from MDR‐TB patients and subjected to semi‐multiplex allele‐specific PCR by using self‐designed primers against the pncA gene. Among 75 samples, 53 samples were subjected to molecular analysis based on purified DNA quantity and quality. The primers produced 250 and 480 bp fragments, indicating the mutations at codon 12 (aspartate to alanine) and codon 85 (leucine to proline) respectively. MDR‐TB was more common in the age group 21–40 years. Fifty‐seven percent of samples (n = 30) were found positive for pncA mutations, whereas 43% of samples (n = 23) showed negative results. Thirteen percent of samples (n = 4) had mutations at codon 12 in which aspartate was converted to alanine, and they produced an amplified product of 480 bp. Eighty‐seven percent of samples (n = 26) had mutations at codon 85 in which leucine was converted to proline and amplified product size was 250 bp. The mutations were simple nucleotide substitutions. The prevalence of mutations in which leucine was substituted by proline was higher than the mutations in which aspartate was substituted by alanine. A high prevalence of substitution mutation (CTG → CCG; leucine to proline) was detected in MDR‐TB cases. Earlier detection of MDR‐TB via an effective molecular diagnostic method can control the MDR tuberculosis spread in the population.

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