Design of primers for identification of honey bee viruses in multiplex-PCR

This paper is devoted to the design of primer oligonucleotide sequences for their use in the genetic identification of Sacbrood virus, Chronic bee paralysis virus, Black queen cell virus and Deformed wing virus using multiplex-PCR. As a result of the bioinformatic analysis, the design of the oligonucleotide primers was performed; the designed primers had similar annealing temperatures (55 C), which makes it possible to indicate each of the viruses under the same PCR conditions. Most of the known strains and isolates of these viruses are amplified with this complex of oligonucleotide primers. Nucleotide sequences of designed primers and a universal positive control allow for the genetic identification of each of the biopathogens under the same PCR conditions at a multiplex format

Isolation of Rabies Virus Glycoprotein Using Three-Phase Extraction and Characteristics of its Antigenic Properties

The aim of the work was to develop an approach to isolation of rabies virus glycoprotein applying threephase extraction and to characterize its antigenic properties.Materials and methods. Infectious activity of the rabies virus (production strain, “Ovine” GNKI) after long-term storage was restored on white BALB/c mice. The strain was used for cultivation on BHK-21 cells; the culture liquid was concentrated applying ultracentrifugation followed by separation by buoyant density in a sucrose gradient, selection of visually opalescent zones, phase concentration, chromatographic separation on an ENrich™ SEC650 column (Bio-Rad, USA) and selection of monomeric fractions with high serological activity according to the results of Western blotting.Results and discussion. We have demonstrated that preliminary mechanical destruction of brain suspension, extraction of the virus-containing material from the cell suspension through successive low-speed and high-speed centrifugation, separation of the sediment produced in sucrose gradient with further phase concentration and chromatographic separation of the precipitate allows to obtain monomeric antigenic preparations with high serological activity. This methodology has made it possible to obtain an antigen, which is rabies virus glycoprotein with a molecular weight of 67 kDa, and two of its isoforms, having molecular weights of 60 and 54 kDa. The described approach can be viewed as an option for isolation of the rabies virus specific antigen when improving laboratory diagnostics techniques. The resulting antigen is a monomeric discrete containing one fraction with a molecu lar weight of 67 kDa. The data obtained corroborate the high specificity of the antigen and its suitability for the design of enzyme immunoassay and immunochromatographic tests, production of specific immunoglobulins, the study of the antigen/antibody interaction, as well as for the assessment of the protective immunity intensity after vaccination

The influence of pollutants on honey bees under conditions of technogenesis of oilproducing regions of the Republic of Tatarstan

Устойчивость медоносных пчел к микозам снижается в районах с наивысшей техногенной нагрузкой. ТМ являются одним из ключевых факторов запускающих механизм снижения резистентности медоносных пчел к микозам. В районах с высокой долей техногенного воздействия пчелы менее устойчивы к аскосферозу и аспергиллезу, значительно меньшую долю составляли меланоз и кандидамикоз. В районах со средней техногенной нагрузкой характер проявления микозов менее выражен, чем в районах с высокой нагрузкой, однако по сравнению с районами с наименьшей степенью нагрузки, в данных районах сохранялась прогрессивная динамика микозов.Resistance of honey bees to mycoses decreases in areas with the highest technogenic loading. HM are one of key factors of the honey bees starting the mechanism of decrease in resistance to mycoses. In areas with a high share of technogenic influence bees are less resistant to ascospherosis and spergillomycosis, a significantly smaller share were melanosis and candidiasis. In areas with average technogenic loading nature of manifestation of mycoses is less expressed, than in areas with high loading, however in comparison with areas with the smallest degree of loading, in these areas progressive dynamics of mycoses remained

Design of primers for identification of honey bee viruses in multiplex-PCR

This paper is devoted to the design of primer oligonucleotide sequences for their use in the genetic identification of Sacbrood virus, Chronic bee paralysis virus, Black queen cell virus and Deformed wing virus using multiplex-PCR. As a result of the bioinformatic analysis, the design of the oligonucleotide primers was performed; the designed primers had similar annealing temperatures (55 C), which makes it possible to indicate each of the viruses under the same PCR conditions. Most of the known strains and isolates of these viruses are amplified with this complex of oligonucleotide primers. Nucleotide sequences of designed primers and a universal positive control allow for the genetic identification of each of the biopathogens under the same PCR conditions at a multiplex format

Lower education level and reduce fish consumption are associated with higher rheumatoid arthritis disease activity: results from the Tatarstan’s women cohort

International audienc

Lower education level and reduce fish consumption are associated with higher rheumatoid arthritis disease activity: results from the Tatarstan’s women cohort.

International audienc

Lower education level and reduce fish consumption are associated with higher rheumatoid arthritis disease activity: results from the Tatarstan’s women cohort

International audienc