P082: Beijing and non-Beijing genotypes detection of mycobacterium tuberculosis by melting curve analyzes

Introduction

Tuberculosis is one of the most important infectious diseases in the world today. Rapid diagnosis of drug resistant Mycobacterium tuberculosis (MTB) is critical to starting of an appropriate treatment and preventing of more spread drug resistant MTB strains.

Objectives

Due to association of Beijing genotype with drug resistance in MTB, we developed a rapid and non-culture method for detection Beijing and non-Beijing MTB in clinical samples.

Methods

We modified Taqman Real time PCR for detection Beijing and non-Beijing genotypes of Mycobacterium tuberculosis to a free probe method in presence of a single dye together with a melting curve analysis. We then performed a blinded screening with both methods on 33 septum samples from treated tuberculosis patients.

Results

We were obtained the same results by both methods. Of the 33 patients, 5 samples were Beijing genotype and 28 were non-Beijing genotype. In free probe method, we were clearly identified a melting peak at 81°C corresponds to non-Beijing and a melting peak at 88°C corresponds to Beijing genotype.

Conclusion

DNA melting curve analysis is a simple and efficient method for the specific detection of amplified products and greatly reduces the cost molecular detection.

Disclosure of interest

None declared.