Usage of chi-Sequence for Differentiation between the Strains of Anthrax Etiological Agent and Closely Related Bacilli

Abstract

Objective of the study was to carry out comparison between the microorganisms appurtenant to six different species of Bacilli genus as regards mobility of the amplicons obtained using the primer that contained chi-sequence of Bacillus subtilis. Therewith, analyzed were 30 Bacillus anthracis strains and closely related bacilli. Cultures grew on L-agar and L-broth at 37 °C. DNA was isolated from vegetative B. anthracis and Bacillus spp. cultures introducing lysozyme and phenol de-proteinizing. In order to perform single-primer PCR constructed was ChiBs primer -5'-CTAGGAGCGGG-3'. Single-primer amplification (30 cycles) was carried out at annealing temperature of 47 °C. Comparative analysis of individual electrophoretic amplicon profiles, obtained by means of ChiBsprimer PCR, allows for identification of genetic intraspecific variability and differentiation of B. anthracis strains from the atypical ones and closely related species of bacilli.

The use of loop-mediated isothermal DNA amplification for the detection and identification of the anthrax pathogen

“…However, the results of these studies were insufficient for unambiguous assignment of these strains to a species. The present study did not yield positive amplification results with primers that targeted the chromosomal loci of the anthrax pathogen, this being in agreement with the published results of single-primer PCR and VNTR typing [34,35] that did not support the assignment of strains 10 and 32 to the B. anthracis species.…”