ISSN 1995-5537
«Біотехнологія» journal V. 5, No. 5, 2012
Р. 65-71, Bibliography 29, Ukrainian.
Universal Decimal classification: 577.2:573.6
DETECTION OF CAUSATIVE AGENT OF ANTHRAX
BY REAL-TIME POLYMERASE CHAIN REACTION
O. Yu. Limanskaya, L. A. Murtazaeva, S. Klee, A. P. Limanskii
Mechnikov Institute of Microbiology and Immunology of National Academy of Medical Sciences of Ukraine, Kharkiv
National Scientific Center «Institute of Experimental and Clinical Veterinary Medicine»
of National Academy of Agrarian Sciences of Ukraine, Kharkiv
Robert Koch-Institut, Berlin, Germany
Correct identification of Bacillus anthracis bacilli remains a challenge for differentiation of the anthrax infectious agent from closely related species B. sereus and B. thuringiensis because of a high homology level of nucleotide sequences with the Bacillus cereus sensu lato instances. In this paper, we have developed molecular genetic sets of primers and probes for real-time polymerase chain reaction with hybridization-fluorescence detection for differentiation of B. anthracis bacilli from В. sereus and B. thuringiensis.
Fragment of ssp gene of chromosomal DNA characterized by hexanucleotide insertion only for B. anthracis isolates was determined as final target for primers and probes. Applying of TaqMan and molecular beacon probes enabled reliable discrimination between B. anthracis bacteria and closely related species B. sereus and B. thuringiensis by real-time polymerase chain reaction. The fluorescence signal for hairpin probes with a molecular beacon format was positive only for B. anthracis strains but for closely related B. sereus and B. thuringiensis species it was negative. Using line TaqMan probes, we registered high-intensity fluorescent signal for all B. anthracis isolates and a signal for B. sereus and B. thuringiensis was much low intensity. The developed approaches could be useful for clinical, epidemiological and epizootiological studies.
Key words: anthrax causative agent, Bacillus anthracis, real-time polymerase chain reaction.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008