RESUMO
The paper presents the results of the development and testing of a molecular biological test system for DNA detection of anthrax pathogen (Bacillus anthracis) by real-time polymerase chain reaction assay. The test system has shown high sensitivity, specificity, and reproducibility of results of analysis, as exemplified by aqueous suspensions of daily agar cultures of Bacillus anthracis strains, related and heterologous species of microorganisms, and clinical materials of experimental animals. There is evidence for the persistence of the basic characteristics of the test system when stored at 22 +/- 2 degrees C for 12 months.
Assuntos
Antraz/diagnóstico , Bacillus anthracis/isolamento & purificação , DNA Bacteriano/análise , Reação em Cadeia da Polimerase/métodos , Animais , Antígenos de Bactérias/genética , Bacillus anthracis/genética , Toxinas Bacterianas/genética , Cricetinae , Humanos , Mesocricetus , Fatores de TempoRESUMO
The genotyping variety of 5 known anthracis vaccine strains using 18 variable loci of the chromosomal localization taken from a microbe culture collection of 48 Research Institute of Ministry of Defense was revealed in the research. The stability of the VNTR-loci was shown to be inherited from the B. anthracis strains with common origin and an opportunity of their gene-identification application. The gene profile of each analyzed vaccine strain using every 18 polymorphic loci was determined and the amplification products were sequenced. The variation of electrophoretic mobility of the amplifiers was found to be caused by the presence of the replication elements with various numbers of copies in their structure.