Comparative study for the rapid detection and genotyping of mtb and rif / inh-resistant mtb mutants with gene flow hybridization and conventional pcr

Authors:Rubina Ghani, Syed Hafeez ul Hassan, Uzma Naseeb, Shamim Mushtaq , Zarghoona Wajid
Int J Biol Med Res. 2017; 8(4): 6121-6124  |  PDF File

Abstract

Tuberculosis (TB) is one of the major infectious causes of morbidity and mortality worldwide. Mycobacteria comprise a diverse group of bacteria that are widespread in nature, some of which cause significant disease in humans. TB is difficult to control due to the time taken for the microbiological diagnosis especially culture on solid media which takes 6–8 weeks. Members of the Mycobacterium tuberculosis complex (MTBC) are the most important human pathogens of the genus Mycobacterium. Traditional methods for detection and identification of mycobacteria include microscopy, culture and phenotypic tests. These methods either lack sensitivity, specificity, or are time consuming. Advances in the field of molecular biology have provided rapid diagnostic tools that have reduced the turnaround times for detecting MTBC and drug resistance in cultures and directly in clinical specimens from weeks to days. The objective of this study was designed to compare Gene Flow with regular PCR and detect the drug resistant mycobacteria in body fluids and tissues besides sputum. The conventional method and PCR detected MTB only in sputum and no sensitivity pattern was observed. In comparison gene flow hybridisation not only detected MTB in sputum, pleural fluid, CSF and tissue samples but it also found the sensitivity patterns. This technology is rapid, cost-effective and the development of the result in an assay is more useful than the single-nested PCR technique for application to diagnosis of a large number of clinical samples.