[The kit of reagents for polymerase chain reaction diagnostic of infections caused by B. pertussis, B. parapertussis and B. bronchiseptica].

The effective treatment of whooping cough directly depends of early diagnostics. The polymerase chain reaction diagnostic is the most perspective diagnostic technique. The kit of reagents is developed to diagnose whooping cough, parapertussis and bronchosepticosis with polymerase chain reaction. The evaluation of its analytical characteristics was carried out. The sensitivity made 1 x 103 of genome equivalents per 1 ml of sample (the sorption technique of DNA extraction was applied) and 5 x 102 of genome equivalents per 1 ml (the precipitation technique of DNA extraction was used). The specificity of test in the framework of analyzed panel of strains and isolates of microorganisms made 100%. The diagnostic sensitivity of analysis exceeded the sensitivity of bacteriological analysis up to 20 times. The application of this kit of reagents permits to detect and to differentiate DNA of agent of whooping cough, parapertussis during one working day already at the beginning of catarrhal period of disease and up to 18th day from the moment of cough appearance. In perspective, this process creates an opportunity to apply timely the specific therapy. The specter of agents of acute respiratory diseases brining on acute prolonged cough in children who were directed to bacteriological analysis to confirm whooping cough is investigated.

[Laboratory diagnostics in evaluation of acute respiratory viral infection morbidity in 2010 - 2011 epidemic season].

AIM: Study of etiological structure of ARVI and evaluation of acute respiratory virus infection morbidity in 2010 - 2011 epidemic season taking into account the data of laboratory diagnostics by method of polymerase chain reaction with hybridization-fluorescent detection. MATERIALS AND METHODS: By using reagent kits produced by Central Research Institute of Epidemiology for the detection of primary causative agents of influenza and ARVI 129 children and 94 adult patients monitored in an outpatient setting as well as 103 children hospitalized due to ARI were examined. RESULTS: Etiological structure of ARVI was studied; proportion of influenza and other actual causative agents of ARVI in monthly dynamics were established. During epidemic rise of influenza (January-March 2011) the proportion of influenza A viruses was 24% (peak in January--31%), the proportion of influenza B viruses--5%, rhinoviruses--9%, metapneumovirus was detected in 6% of cases, parainfluenza viruses (1 - 4 type) and adenovirses--4% each, coronaviruses--in 3%, respiratory syncytial virus--in 2%, bocavirus--in 1% of the studied samples. In influenza structure A/H1N1pdm2009 virus, its proportion was 70%, influenza virus B (26.9%), influenza virus A/H3N2 (2.6%) predominated. Indexes for monthly morbidity caused by each of the ARVI causative agents were calculated. CONCLUSION: The proposed approach allowed to evaluate ARVI morbidity taking into account laboratory-confirmed etiological factors. A 5 time increase in ARI morbidity in adults in February 2011 was shown to be mostly due to an increase in influenza A morbidity as well as involvement of influenza B virus, metapneumoviruses, coronaviruses, parainfluenza viruses and rhinoviruses into the epidemic process. Increase of morbidity of children by 1.4 times was also seen during activization of influenza viruses and metapneumovirus. The analysis of monitoring results allowed to prognose increase of respiratory-syncytial viral infection epidemic activity from September 2011 to February 2012.

[Molecular characterization of pandemic influenza viruses A/H1N1(sw2009) isolated in Russia in 2009-2010].

AIM: Assessment of genetic diversity of influenza virus A/H1N1 (sw2009) variants circulated in Russia, study of virus' pathogenicity in humans and potential resistance to antiviral drugs. MATERIALS AND METHODS: Sequencing of PCR-fragments of genome of influenza viruses isolated from clinical and autopsy samples of 436 patients. Four full genome sequences of influenza viruses A/H1N1 (sw2009) were obtained. Phylogenetic analysis was performed. RESULTS: High degree of homology (98.9-100%) was found among influenza A/H1N1(sw2009) viruses in HA and NA genes as well as in their aminoacid sequences (1.3 and 1.4% respectively). Differences in other proteins did not exceed 1.1%. Diversity was found in position 222 of receptor-binding locus of HA and single amino acid polymorphism--in several internal proteins. Known mutations determining resistance to Tamiflu and Arbidol were not detected. All viruses were resistant to remantadine. Molecular markers of high pathogenicity were not found. CONCLUSION: High homology of influenza viruses determines low level of antigenic differences although in populations of viruses there are variants with different levels of adaptation to human organism and different affinity to receptors of upper and lower respiratory tract that can determine their different transmissibility.

[Pandemic influenza A/H1N1 (sw2009) in Russia: epidemiology, diagnosis, clinical picture, and treatment].

AIM: To study the epidemiological and clinical features of the 2009-2010 pandemic influenza in Russia. SUBJECTS AND METHODS: Materials from 874 patients, including postmortem samples from 287 subjects, were examined applying the AmpliSens Influenza virus A/H1-swine-FL PCR kit designed and produced by the Central Research Institute of Epidemiology. The clinical and postmortem characteristics of 68 patients who had died from influenza A/H1N1 (sw2009) were analyzed in detail. RESULTS: The cause of deaths was primary virus pneumonia in most cases. The major manifestation of viral pathogenicity was impaired microcirculation leading to hemorrhage. No mutations conferring resistance to oseltamivir and arbidol were found. All A/H1N1swl viruses had genetic markers of remantadin resistance. CONCLUSION: The reagent kits developed by the Central Research Institute of Epidemiology proved to be effective. It is necessary to set up PCR laboratories that differentially diagnose influenza and acute respiratory viral infections in health care facilities in order to make early laboratory diagnosis of influenza and to timely perform its specific therapy.