RESUMO
Diagnostic properties of new monoclonal antibodies (MAbs) to hexon adenovirus antigen (AB) monoclonal ELISA kit for early diagnosis of adenoviral infection were tested. Developed ELISA kit and FITC-conjugate of new monocional antibodies for immunofluorescent analysis were used for detection of different types of adenoviruses in clinical materials. The availability of their use in clinical and epidemiological practice was validated.
Assuntos
Infecções por Adenoviridae/diagnóstico , Adenovírus Humanos/imunologia , Anticorpos Monoclonais/biossíntese , Anticorpos Antivirais/sangue , Antígenos Virais/imunologia , Infecções por Adenoviridae/sangue , Infecções por Adenoviridae/imunologia , Infecções por Adenoviridae/virologia , Adenovírus Humanos/classificação , Adenovírus Humanos/genética , Animais , Anticorpos Monoclonais/química , Anticorpos Monoclonais/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/normas , Fluoresceína-5-Isotiocianato/química , Humanos , Imunoconjugados/química , Camundongos , Kit de Reagentes para Diagnóstico , Sensibilidade e EspecificidadeRESUMO
In the surveillance of rubella in the northwest region of Russia samples of nasopharyngeal swabs from 37 patients with rubella, which were treated in the 442nd district military hospital named after Z.P. Solovyov in autumn 2007 were screened for the rubella virus using RK-13 cell line, 22 strains of rubella virus were isolated. Gene sequencing of E1 region of rubella virus isolates was carried out. Rubella virus strains isolated in St. Petersburg during the 2007 outbreak belonged to rubella virus genotype 1E. The morphogenesis of RK-13 cells with formation of replication complexes and enveloped virions of rubella virus was shown.
Assuntos
Células Epiteliais/virologia , Nasofaringe/patologia , Vírus da Rubéola/genética , Rubéola (Sarampo Alemão)/diagnóstico , Proteínas do Envelope Viral/genética , Adolescente , Adulto , Animais , Linhagem Celular , Surtos de Doenças , Células Epiteliais/patologia , Hospitais Militares , Humanos , Masculino , Nasofaringe/virologia , Filogenia , Reação em Cadeia da Polimerase , Coelhos , Rubéola (Sarampo Alemão)/epidemiologia , Vírus da Rubéola/classificação , Vírus da Rubéola/isolamento & purificação , Federação Russa/epidemiologia , Proteínas do Envelope Viral/metabolismo , Adulto JovemRESUMO
The results of molecular genetic analysis of more than 280 strains of influenza A virus subtypes H1N1 and H3N2 circulating in Russia in 2006-2012 are presented. The genetic changes underlying the evolution of the virus strains and sensitivity to antiviral drugs were analyzed. Significant changes in the genetic structure of influenza A viruses circulating in the Russian Federation and their phylogenetic affiliation are shown to occur within the studied period. The studies identifying codons under the positive selection in silico in the genes encoding surface proteins of the influenza virus were demonstrated to be efficient for the analysis of the antigenic drift and direction of evolutionary variability of the influenza viruses.
Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza , Vírus da Influenza A , Influenza Humana , Filogenia , Evolução Molecular , Deriva Genética , Variação Genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/classificação , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Humanos , Vírus da Influenza A/classificação , Vírus da Influenza A/genética , Influenza Humana/classificação , Influenza Humana/genética , Influenza Humana/virologia , Federação RussaRESUMO
The analysis of 1558 clinical samples revealed influenza virus A(H1N1v) RNA in 339 patients with influenza and 163 fatal cases,which was made in May to December 2009. Data on the antigenic properties of more than 250 of pandemic virus strains isolated at the Research Institute of Influenza and the molecular genetic characteristics of 31 strains are presented. All the test isolates were found to have the S203 substitution in hemagglutinin, which was characteristic of one of 5 minor genome A(H1N1v) virus variants found in the United States and Mexico in 2009. All the test strains contain the S31N substitution in the M2 protein, which determines viral resistance to adamantine, and have no H275Y substitution in neuraminidase, which determines oseltamivir resistance. The substitution of amino acid residue of Asp to Gly at position 222 of HA was found in 8 (73%) of 11 isolates from postmortem lung and trachea samples and in 2 (10%) of 20 isolates from nasopharyngeal swabs. The determination of the pathogenic role of this substitution calls for further investigations.
Assuntos
Hemaglutininas/genética , Vírus da Influenza A Subtipo H1N1/genética , Influenza Humana/virologia , Neuraminidase/genética , Vírus Reordenados/genética , Proteínas da Matriz Viral/genética , Adolescente , Adulto , Idoso , Amantadina/análogos & derivados , Amantadina/farmacologia , Amantadina/uso terapêutico , Substituição de Aminoácidos/efeitos dos fármacos , Animais , Antivirais/farmacologia , Antivirais/uso terapêutico , Embrião de Galinha , Criança , Pré-Escolar , Farmacorresistência Viral/efeitos dos fármacos , Farmacorresistência Viral/genética , Humanos , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Influenza Humana/diagnóstico , Influenza Humana/tratamento farmacológico , Influenza Humana/mortalidade , Pulmão/virologia , México , Pessoa de Meia-Idade , Mortalidade , Nasofaringe/virologia , Oseltamivir/farmacologia , Oseltamivir/uso terapêutico , Pandemias , Filogenia , Vírus Reordenados/efeitos dos fármacos , Vírus Reordenados/isolamento & purificação , Federação Russa , Traqueia/virologia , Estados Unidos , Proteínas Virais/genética , Adulto JovemRESUMO
The paper is based upon the results of clinic-pathological and virological correlations in 29 lethal cases of influenza in Saint-Petersburg and Leningrad region during the epidemics 2009/2010. Immunohistochemical analysis of lungs, heart and brain using monoclonal sera to HA and HP proteins of influenza virus, virological and morphological analysis of experimental influenza in mice infected by A/WSN/33 (HIN1) and A/California/07/09 (H1N1) viruses had been carried out. In the majority of investigated strains was proved the amino acid mutation with replacement D222G. The replication of virus was demonstrated at the late stages of diseases, but the desquamation of respiratory epithelium and cytoproliferative weren't found out. Besides the "influenza cells", previously described by A. V Zinserling the cells with enlarge light nuclei were observed. Patients with influenza died from respiratory distress syndrome with minimal bacterial infection. We've established that H1N1 virus not only damages the cells of respiratory epithelium and alveolar macrophages but it can injure endothelium of different organs and neuroglia. The questions which have to be discussed are listed.
Assuntos
Encéfalo/patologia , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Influenza Humana , Pulmão/patologia , Miocárdio/patologia , Adolescente , Adulto , Animais , Autopsia , Encéfalo/virologia , Feminino , Coração/virologia , Humanos , Imuno-Histoquímica , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N1/fisiologia , Influenza Humana/mortalidade , Influenza Humana/patologia , Influenza Humana/virologia , Pulmão/virologia , Masculino , Camundongos , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Síndrome do Desconforto Respiratório/etiologia , Síndrome do Desconforto Respiratório/mortalidade , Síndrome do Desconforto Respiratório/patologia , Síndrome do Desconforto Respiratório/virologia , Federação Russa/epidemiologia , Replicação Viral , Adulto JovemRESUMO
Russian enzyme immunoassay (EIA) test systems (EATS) and foreign analogues (DSL and BCM-Diagnostics, USA) were compared in the detection of human serum rubella virus (RV) antibodies. Analysis of RV IgM levels ascertained a greater agreement of quality indices for the EATS "EKOlab-Krasnukha-IgM" than for the system "VectoRubella-IgM". As compared with the USA reference systems, the sensitivity, specificity, and total agreement of the data obtained by the EATS "EKOlab" were 100, 97.5, and 97.7%, respectively; those were 88, 84.4%, and 85.4% for "VectoRubella-IgM". In healthy individuals, strictly seropositive cases of IgM detection, revealed by the EATS "VectoRubella-IgM" and unconfirmed by the results obtained by the EATS "BCM-Diagnostics-IgM" (8%) are most likely to be regarded as false-positive due to the presence of serum rheumatoid factor (RF). The principle of indirect EIA used in the system "VectoRubella-IgM" makes it impossible to rule out the impact of RF on test results. The EATS "EKOlab-Krasnukha-IgM" and the systems made in the USA apply the principle of EIA with IgM involvement that, unlike indirect EIA, minimize to have nonspecific results. All three analyzed Russian EATS "EKOlab-Krasnukha-IgG", "Krasnukha-screening", and "VectoRubella-IgG" in the detection of RV IgG show rather high diagnostic parameters as compared with the systems made in the USA. The important additional advantage of the EATS "EKOlab-Krasnukha-IgG" over other Russian systems is that the kit has reference serum with the known content of RV IgG. This allows one to give results in absolute units and to standardize the calculations of some independent experiments. The performed study suggests that out of the Russian test systems, EATS "EKOlab-Krasnukha-IgM" and "EKOlab-Krasnukha-IgG" should be preferred.
Assuntos
Anticorpos Antivirais/sangue , Técnicas Imunoenzimáticas/normas , Vírus da Rubéola/imunologia , Adolescente , Adulto , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Kit de Reagentes para Diagnóstico , Rubéola (Sarampo Alemão)/diagnóstico , Rubéola (Sarampo Alemão)/virologia , Sensibilidade e Especificidade , Testes Sorológicos , Adulto JovemRESUMO
AIM: To reveal features of rubella epidemic process during start of mass immunization and to determine rubella virus genotypes circulating in Saint-Petersburg. MATERIALS AND METHODS: Official data on rubella morbidity during 1995-2007 and number of vaccinated against rubella children and adults were used in this study. During 2006-2008 males aged 17-20 years with rubella diagnosis were eligible for laboratory test on rubella. Nasopharyngeal swabs and blood specimenswere tested by PCR and virus isolation on cell culture (PK13). Genotyping of isolates was performed on the basis of 600 nucleotide sequence of E1 gene from 8731 to 9653 n.p. RESULTS: It was shown that mass vaccination of children and young women against rubella during 4 years resulted in 3-fold drop of rubella incidence inwhole population, which diminishes the probability of infection in pregnant women and born of children with congenital rubella syndrome. In age structure of rubella morbidity the proportion of children aged 3-6 and 7-14 years decreased by 1.5-fold. Epidemic process loss the features of autoregulating system (periodicity and seasonal incidence peaks). Results of genotyping showed that isolates belonged to genotype 1E. High degree of homology (97.7-99.6%) to isolates from Barnaul and Belorussia was demonstrated. CONCLUSION: Issues on isolates' origin and success of measures on elimination of endemic rubella could be resolved by further studies on isolation and genotyping of rubella virus strains in Saint-Petersburg and North-East region of Russian Federation in the whole.