Abstract

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a deadly infectious disease. India contributes to one-third of the global TB burden. However, no studies have been carried out in the Telangana (Hyderabad) population using real-time polymerase chain reaction (RT-PCR). Therefore, the current study evaluated the role of RT-PCR as a rapid and non-invasive test to diagnose TB by testing for pulmonary tuberculosis (PTB) and extrapulmonary tuberculosis (EPTB). This hospital-based study examined 1670 samples (900 EPTB; 770 PTB) comprising tissue (n = 537), peritoneal fluid (n = 420), sputum (n = 166), bronchial fluid (n = 126), cerebrospinal fluid (n = 145), ascetic fluid (n = 76), sputum pus (n = 78), urine (n = 79), and bronchoalveolar fluid (n = 43) samples. DNA from samples was separated using specific isolation kits and subjected to RT-PCR. In this study, we enrolled 1670 subjects and categorized 54.4% as females and 45.6% as males. The collected samples were categorized as 48.5% of fluid samples, followed by tissue (32.2%), sputum (9.9%), urine (4.7%), and pus-swab (4.6%). RT-PCR analysis revealed that 4.7% patients were positive for Mtb. Our results revealed that 61% of the affected patients were male and 39% were female. Among the specimen types, tissue samples gave the highest proportion of positive results (36.3%). The results showed that RT-PCR should be implemented and given top priority in TB diagnosis to save time and facilitate a definitive diagnosis. Tissue samples are highly recommended to screen the Mtb through the technique RTPCR. Future studies should extend the technique to the global population and exome sequencing analysis should be performed to identify TB risk markers.

Highlights

  • Tuberculosis (TB), caused by acid-fast aerobic bacteria comprising the Mycobacterium tuberculosis (Mtb) complex, is a contagious infection that generally affects the lungs

  • The current study evaluated the role of real-time polymerase chain reaction (RT-PCR) as a rapid and non-invasive test to diagnose TB by testing for pulmonary tuberculosis (PTB) and extrapulmonary tuberculosis (EPTB)

  • The current study aimed to evaluate the role of real-time polymerase chain reaction (RT-PCR) to test PTB and EPTB specimens as a rapid and non-invasive test for the fast diagnosis of TB

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Summary

Introduction

Tuberculosis (TB), caused by acid-fast aerobic bacteria comprising the Mycobacterium tuberculosis (Mtb) complex, is a contagious infection that generally affects the lungs. In cases of suspected EPTB, rapid and accurate laboratory diagnosis is important, because traditional techniques of detecting acid-fast bacilli have limitations. Molecular techniques have proven very successful as diagnostic tools These molecular typing techniques target polymorphic genetic sequences, but interrogate less than 1% of the genome; they have an intrinsically restricted discriminatory power. This limitation could be overcome by the application of next-generation whole-genome sequencing (WGS) for genome-based epidemiology. The current study aimed to evaluate the role of real-time polymerase chain reaction (RT-PCR) to test PTB and EPTB specimens as a rapid and non-invasive test for the fast diagnosis of TB

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