isolates from kermanshah province submitted at national animal tuberculosis reference laboratory at razi institute

Rogheih Moghaddam,1 Nader mosanari,2 Ardeshir hesamour,3,* -,4 -,5

1. Razi institute
2. Razi institute

Abstract


Introduction

Tuberculosis is a zoonotic which has long been recognized and is still one of the most life-threatening diseases. tuberculosis bacterium is the second infectious agent that causes mortality in humans alone. this study aims at determining the identity of mycobacterium tuberculosis complex species from each other for rapid identification of species and successful treatment of the disease.

Methods

first fifty isolates were grown on lowenstein jensen medium (ljp and ljg). the colonies were observed by light microscopy com¬bined with a zn stain smear and fluorescent microscopy with fluorochrome staining.then genomic dna was extracted from the bacterial isolates by boiling and the isolates identified to genus level by pcr-16srrna. pcr-is6110 was used to identify the isolates belonging to mycobacterium tuberculosis complex while mycobacterium species were identified based on pcr-rd typing (,rd9 and rd12). finally to identify&confirm species specially species of m.tuberclusis, m.bovis,m.bovis bcg. we usedpcr-rflp using oxy-r genes

Results

Result: : it was proven that all cases of 50 isolates sent were m.tuberclusis

Conclusion

It is worth mentioning that most tuberculosis cases are identified on the basis of acid-fast bacilli detection, and antibiotic therapy is immediately initiated subsequently. moreover, it should be noted that some of these acid-fast positive cases might not be of genus mycobacterium, and thus, the antibiotics prescribed might threaten the health of the patients. additionally, if the identified bacilli are not within mtb complex, the drug therapy would differ. however, mycobacterium bovis, which is a member of mtb complex and is resistant to pyrazinamide, requires exact strain identification. in conclusions we suggest that individual isolates should be identified by rd typing& pcr-rflp of oxy-r methods, which has discriminatory power for accurate species identification of mycobacterium species.

Keywords

M.tuberclusis,pcr-rd typing ,pcr-16srrna, pcr-is6110