Molecular identification of M. bovis BCG by Multiplex PCR

Abd El-Tawab, Ashraf A.; El-Hofy, Fatma I.; Nasr, Essam A.; Sriranganathan, Nammalwar; Soliman, Enas A.

doi:10.21608/bvmj.2016.31236

Molecular identification of M. bovis BCG by Multiplex PCR

Document Type : Original Article

Authors

¹ Department of Bacteriology, Immunology and Mycology Fac. Vet. Med. Benha University, Egypt

² Department of diagnostic bacteriology. Veterinary serums and vaccines research Institute, Abbasia, Cairo, Egypt

³ Department of Biomedical Sciences & Pathobiology, Virginia-Maryland College of Veterinary Medicine, USA

10.21608/bvmj.2016.31236

Abstract

Mycobacterium bovis (M. bovis) BCG belongs to Mycobacterium tuberculosis complex (MTBC), highly related
organisms, which are 99.9 % similar at nucleotide level and phenotypically similar. Its differentiation from other members
of MTBC by conventional methods is laborious and time consuming. PCR provides a rapid alternative method for
differentiation between members of MTBC. It depends on identification of region of difference (RD) which have been
lost during attenuation of M. bovis. Two different BCG strains (from two sources) were confirmed as a member of M.
tuberculosis (MTB) complex (MTC) and as BCG strains by PCR using primers to a region of the 16S rRNA gene that is
conserved in all mycobacteria and region of difference (RD1, RD4, RD9 and RD12) respectively. DNA of Mycobacterium
tuberculosis was used as a control to compare with BCG strains. The results showed that 16S rRNA gene was present in
all tested strains, while the RD1, RD4, RD9 and RD12 were absent only in BCG strains

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