RESUMO
The full-scaled agglutinating immunoassay is commonly applied to detect content of antibodies to cholera agent Vibrio cholerae human in blood serum under application of serological diagnostic. The time of analysis implementation amounts to 18 hours. To shorten time of detection of antibodies a biological microchip (biochip) was developed. The biochip represents an activated slide with immobilized corpuscle and soluble antigen cholera agent (O-antigens, cholera toxin). The experimental work resulted in development of scheme of biochip and selection of optimal conditions of sorption and implementation of immunologic analysis using biochip. The application of biochip facilitated to detect specific antibodies to antigens of cholera agent in commercial experimental animal serums and blood serums of ill patients. The time of analysis implementation amounted to 2-3 hours. The results are substantiated by bacteriological and serological methods.
Assuntos
Anticorpos Antibacterianos/sangue , Cólera/sangue , Análise Serial de Proteínas/instrumentação , Vibrio cholerae/isolamento & purificação , Animais , Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/química , Antígenos de Bactérias/imunologia , Cólera/imunologia , Cólera/microbiologia , Toxina da Cólera/química , Toxina da Cólera/imunologia , Humanos , Vibrio cholerae/imunologiaRESUMO
AIM: Detection of circulation of West Nile virus (WNV) on the territory of Saratov Region and prerequisites for formation of natural focus of West Nile fever (WNF), determination of the role of WNV in infectious pathology on the territory of the region. MATERIALS AND METHODS: of organs of small mammals, birds, blood-sucking arthropods for the presence of WNV markers (antigens and/or RNA) were studied. Clinical material from patients with symptoms not excluding WNF was studied. Donor blood sera samples were analyzed with the aim of detection of immune layer against WNV in population of Saratov Region. RESULTS: In 2010 WNV antigens were detected by EIA in 12 samples (7.1%) of mammal organ suspensions. In 2012 by using RT-PCR and EIA, markers of WNV were detected in 6 samples of bird brain suspensions (6.3%) and 1 sample of mammal organ suspension. Immune layer of population against WNV was 4% in 2011, 2.8% in 2012. In 2012 in 11 of 27 examined patients IgM against WNV in diagnostic titers and/or serconversion of IgG in paired sera were detected. In addition in 5 individuals virus RNA was detected in blood. Based on clinical, laboratory data and epidemiologic anamnesis 11 patients were diagnosed with WNF. CONCLUSION: The results obtained give evidence on the circulation of WNV on the territory of Saratov Region in 2010 - 2012. With the development of complications of WNF epidemiologic situation in 2012 an expansion of WNV areal onto the territory of the region took place and the process of formation of WNF natural foci is ongoing.
Assuntos
Febre do Nilo Ocidental , Vírus do Nilo Ocidental , Animais , Anticorpos Antivirais/sangue , Aves/sangue , Aves/virologia , Humanos , Imunoglobulina M/sangue , Federação Russa/epidemiologia , Febre do Nilo Ocidental/sangue , Febre do Nilo Ocidental/epidemiologia , Febre do Nilo Ocidental/transmissãoRESUMO
A study of the structural and regulatory genes, determining rhamnose fermentation, that are located in the rha locus of the chromosome of Yersinia pestis main and non-main subspecies and of Yersinia pseudotuberculosis of serogroups I-III was performed. The nucleotide sequence of Y. pestis main subspecies differs substantially from those of non-main subspecies and Y. pseudotuberculosis by the presence of a nucleotide substitution in 671 bp location of rhaS gene resulting presumably in the Y. pestis non-main subsp inability to utilize rhamnose. This results in incapability of Y. pestis non-main subspecies to utilize rhamnose. Other nucleotide substitutions found in Y. pestis non-main subspecies strains influence only upon expression efficiency of this diagnostic criterion.