Detection of Non-Tuberculosis Mycobacteria by Real Time PCR

Abstract

Background:: Identification of non-tuberculosis mycobacteria by culture and phenotypic description is commonly used; however, it takes 4 to 6 weeks or even a longer time for slow growing species as well as for identification of some species that may be missed by biochemical characteristics methods. This study aimed to evaluate Real Time PCR for Detection of NTM by Amplification of Internal Transcribed Spacer (ITS) and 16S rRNA. Methods:: In our investigation, using Real Time PCR and two pairs of unique primers targeted to ITS and 16S rRNA genes as well as Beta-actin as an internal control, Non tuberculosis mycobacteria species were detected. Results:: Real time PCR was performed on the prepared dilutions. In addition, the threshold of sensitivity in this study was 10pg. To test the specificity, the genome of several bacteria responsible for respiratory infections was used, in which only the test response related to the non-tuberculosis mycobacterium genome and internal control was positive. Conclusion:: In this research, an effective and up-to-date Real Time PCR method was used to design a diagnostic kit from all aspects. To avoid any error or mistake and to minimize the false results, internal control was used. The ability to design diagnostic kits allows us to increase efficiency, minimize mistakes, and save a considerable amount of time and cost.