ABSTRACT
An enzyme immunoassay-based test system for Y. pestis V antigen detection was developed. The specificity and sensitivity of this system met the requirements for medical immunobiological preparations for the identification of causative agents of highly fatal diseases. The sensitivity of the test system was assessed, and its high specificity was also demonstrated: the test system did not detect bacterial cells of closely related (four Y. pseudotuberculosis strains) and heterologous microorganism strains. The test system developed was able to detect the V antigen at concentrations as low as 2.0 ng/mL in cells of nine experimental Y. pestis cultures. The obtained preparation can be recommended for use in laboratory diagnostics of plaque.
Subject(s)
Antibodies, Monoclonal/chemistry , Antigens, Bacterial/analysis , Immunoenzyme Techniques/standards , Pore Forming Cytotoxic Proteins/analysis , Yersinia pestis/isolation & purification , Animals , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/isolation & purification , Antibody Specificity , Antigens, Bacterial/immunology , Humans , Hybridomas/immunology , Immunoblotting , Limit of Detection , Mice , Mice, Inbred BALB C , Plague/diagnosis , Plague/microbiology , Pore Forming Cytotoxic Proteins/immunology , Yersinia pestis/chemistry , Yersinia pestis/immunology , Yersinia pseudotuberculosis/chemistry , Yersinia pseudotuberculosis/immunology , Yersinia pseudotuberculosis/isolation & purificationABSTRACT
Molecular mechanisms of the Yersinia pestis pathogenicity and peculiarities of maturation of specific immunity to plague are reviewed. The history and modern state of the plague vaccine development are described. Special attention is focused on the prospects in the area of the plague vaccine development. The possible approaches to improvement of vaccine preparations are discussed.
Subject(s)
Plague Vaccine , Plague , Yersinia pestis , Animals , Humans , Plague/epidemiology , Plague/genetics , Plague/immunology , Plague/metabolism , Plague/prevention & control , Plague Vaccine/immunology , Plague Vaccine/therapeutic use , Yersinia pestis/genetics , Yersinia pestis/immunology , Yersinia pestis/metabolism , Yersinia pestis/pathogenicityABSTRACT
57 Y pestis bv. caucasica strains were assayed using molecular typing. The results of these assays indicated the presence within this biovar of the three separate clonal clusters and necessity of detachment of the Leninakan mountain mesofocus (subfocus) from the structure of Transcaucasian-highland focus into self-supporting one, as well as inclusion of a part of the Pre-Araks low-mountain natural plague focus in the capacity of the subfocus along with Pre-Sevan mountain and Zanzegur-Karabakh mountain subfoci into the structure of Transcaucasian-highland focus. It was shown that the strains circulating in the East-Caucasian highland plague focus were the most ancient branch of bv. caucasica or even of the entire Y pestis phylogenetic tree.
Subject(s)
Phylogeography , Plague , Yersinia pestis/genetics , Animals , Antigens, Bacterial/genetics , Arvicolinae/microbiology , Bacterial Proteins/genetics , Humans , Minisatellite Repeats/genetics , Plague/genetics , Plague/microbiology , Pore Forming Cytotoxic Proteins/genetics , Serine Endopeptidases/genetics , Yersinia pestis/classificationABSTRACT
The main shortcoming of the modem live and killed vaccines based on gram-negative bacterial strains is their ability to cause adverse reactions. The majority of the adverse reactions are associated with the effect of biological activity of lipopolysaccharide. The report covers the problems concerned with biogenesis of the lipid A, lipopolysaccharide structural component, responsible for its endotoxic activity, as well as with genes determining lipid A synthesis. The special attention is paid to gene-engineering technique for reduction of adverse reactions of vaccine strains that are based on the knock-out mutagenesis of genes waaM and/or waaN responsible for addition of lauroyl and myristoyl residues to the distal glucosamine unit lipid A, generating acyloxyacyl moieties.