Assessment of the Viability of Mycobacterium Leprae from Soil Samples using Real Time RT- PCR Targeting 16S rRNA Gene

Investigating the Viability of Mycobacterium leprae in Soil using Real Time RT-PCR Analysis

Abstract

Leprosy is an infectious and communicable disease caused by Mycobacterium leprae, which affects the skin and nerves. Leprosy has ceased to be a public health problem world-wide, after the successful implementation of effective chemotherapy and use of control measures. The present retrospective study was the part of MSc Biotechnology program and was conducted to assess the viability of Mycobacterium leprae from soil samples by Real time RT- PCR targeting 16S rRNA gene. In this retrospective study 40 soil specimens from 20 villages from Ghatampur region were taken and viability was assessed by Real time RT-PCR analysis targeting 16S rRNA gene region. RNA of M. leprae extracted and identified using specific M. leprae primers. PCR amplification was done and time period RT-PCR was accustomed observe viable M. leprae. Viable Mycobacterium leprae were detected by real time RT-PCR analysis targeting 16S rRNA gene region from 13out of 40 (32.5) soil samples. The copy number of M.leprae estimated varied from 2.9x101 to 3.05x103 100mg soil in the positive samples using the present protocol. Real time PCR targeting 16S rRNA appears to a promising approach for studying the presence of live M.leprae in the environment. This study therefore provides valuable data of presence of viable M. leprae in soil specimens, which might be of use in work the transmission dynamics in Hansen's disease.