Strain of influenza virus a/17/mallard/netherlands/00/95(h7n3) for production of live and inactivated influenza vaccines. RU patent 2507256.
 Species-specific bacteriophage strain having lytic activity in relation to staphylococcus aureus, including multi-drug resistant strains / 2503716Strain is deposited under number Ph 62 in GKPM-Obolensk collection. Strain can be used for development of complex medical and disinfective preparations against infections caused with Staphylococcus aureus, as well as preparations for sanitation of food products against Staphylococcus aureus. |
 Extracted polynucleotide molecule coding torque teno virus, rna molecule and expression vector / 2502801Group of inventions refers to biotechnology and deals with new nucleotide sequences of Torque teno virus (TTV) and vectors containing such sequences. Extracted polynucleotide molecule contains polynucleotide sequence chosen from the group consisting of SEQ ID NO:4, sequence complementary to SEQ ID NO:4 and polynucleotide that is at least 95% identical to SEQ ID NO:4. |
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Method for preparing cattle viral diarrhoea vaccine / 2502522 Invention refers to veterinary virology, microbiology and biotechnology and may be used in developing the agents for specific prevention, particularly for preparing cattle viral diarrhoea vaccine. To improve the quality of an end product by using an oxidant solution as an inactivating agent prepared by the electrolysis of sodium chloride solution, as well as to activate the method for preparing the cattle viral diarrhoea vaccine involving preparing a virus-containing material of the cattle viral diarrhoea vaccine strain, infecting a continuous cell culture with the virus-containing material, culturing the cattle viral diarrhoea virus, collecting a virus-containing fluid, inactivating and preparing the end product in the liquid form, with the cattle viral diarrhoea virus being inactivated by the oxidant solution prepared by electrolysis of 10.0-20.0% sodium chloride with the electrolysis carried out to pH 7.0-8.0, oxidant concentration 0.7-0.9% and redox potential +1000±50 mV; the processing is one-staged with the active chlorine content Cax=400-600 mg/l for 60-70 min with inactivating agent consumption 4.5-5.0 cm3 per 0.8-1.0 l of the virus-containing fluid. |
 Strain of bacteriophage escherichia coli ecd4, having lytic activity in respect to bacteria escherichia coli of serotype o104:h4 / 2496874Invention relates to a strain of the bacteriophage Escherichia coli ECD4. The strain of the bacteriophage Escherichia coli ECD4 is isolated from faeces of broiler chickens on the culture of the bacteria of the strain Escherichia coli O104.H4 RK1No.112027 and is deposited in the State Collection of Pathogenic Microorganisms and Cell Cultures "GKPM-Obolensk" under the number Ph63. The proposed strain of the bacteriophage has lytic activity in respect to the bacteria of the strain Escherichia coli O104:H4 RKINo.112027, lyses Escherichia of the serotype 0157:H7, does not suppress growth of cells Escherichia coli M-17, has lytic activity in respect to several other clinically significant serotypes of Escherichia, and also to several types of Shigella. The strain of the bacteriophage Escherichia coli ECD4 propagates on the laboratory non-pathogenic strain Escherichia coli K-12 C600F. |
 Strain of enterovirus coxsackie b6 selectively infecting and lysing tumorous human cells in vitro / 2496873Described strain is produced by performance of a series of adaptation passages of a parent strain of the virus ZhEV-15 Coxsackie B6 on the culture of cells HEK293 highly sensitive to this virus and the neoplastic cell line C33A (HPV-negative carcinoma of human uterine neck), with production of a new strain ZhEV-15L of the virus Coxsackie B6. The strain selectively infects and lyses tumorous human cells and has a fragment of a genome sequence represented in the dwg. 2, being its marker criterion. The strain is deposited in the State Collection of the Federal Service for Oversight of Consumer Protection and Welfare of agents of viral infections and rickettsial diseases of the Federal Budget Institution of Science State Science Centre of Virusology and Biotechnology "Vector" under the registration number V-576. |
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Strain of flu virus a/common gull/altai/804/2011/h16no-subtype for production of antigene-containing preparation, polyclonal serum and application as control reference-sample when assessing specificity of test-systems based on polymerase chain reaction / 2496872 Invention relates to the field of microbiology and relates to the strain of flu virus of H16N3-subtype. The strain of bird flu virus A/common gull/Altai/804/2011/H16N3-subtype is described, deposited in the State Collection of Viral Infectons and Rickettsial diseases of the Federal Budget Institution of Science State Science Centre of Virusology and Biotechnology "Vector" under the registration number V-583. The strain was isolated from a common gull (Larus canus). The strain may be used to produce a polyclonal serum to determine affinity of the flu virus to H16-subtype, and also may be used as a control reference sample when assessing specificity of test systems on the basis of PCR. |
 Genes coding major capsid protein l1 of human papilloma virus, and using them / 2494106Invention refers to biotechnology and virology. The present invention discloses a codon-optimised gene coding the major capsid protein L1 of human papilloma virus which is able for effective expression of the major capsid protein L1 of human papilloma virus after transducing into a yeast cell. What is also described is an immunogenic macromolecule which is preferentially formed by the expression of the above codon-optimising gene coding the major capsid protein L1 of human papilloma virus in the yeast cell. What is also disclosed is using the above immunogenic macromolecule and composition containing the above immunogenic macromolecule. |
 Methods and compositions for immunisation of pigs against porcine circovirus / 2493254Nucleic acids are described, which code genomes of two new strains of porcine circovirus of type 2B. These two new strains of porcine circovirus may be used to produce a vaccine or an immunogenic composition for immunisation of pigs against porcine multisystemic wasting syndrome (PMWS). |
 Attenuated strain "kem-7" of fowlpox virus for manufacturing of preparations of specific prevention and diagnostics of fowl pox / 2493253New strain "KEM-7" of the fowlpox virus is produced by multiple passages of mother virus on developing chicken embryos. The strain is deposited into the collection of FSBI VGNKI under the registration number (reference) - strain "KEM-7" - DEP of fowlpox virus. The strain is genetically authentic by the sequence of nucleotides of the gene 241ext. It is adapted to the organism of developing chicken embryos. It is immunogenic, moderately reactogenic and harmless for birds in case of an intradermal injection. It preserves the attenuated phenotype during passages both in the cultivation system and on a naturally receptive bird. |
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Rubella virus strain "orlov" (u) for obtaining medicinal immuno-biological preparations (mibp) / 2492235 Strain was deposited under number V-1 in the collection "SCPM-Obolensk". The strain may be used to produce monovalent rubella vaccine, associated combined vaccines comprising the vaccine strain of rubella virus, as well as for production of diagnostic products. |
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Method for specific prevention of herd equine rhinopneumonia, salmonella abortion and strangles with using associated vaccine / 2506093 Invention relates to veterinary science. A method involves immunising with using an associated inactivated vaccine prepared of equal portion of a salmonella abortion vaccine prepared of the bacterial strain Sal. abortus equi BN-12, an inactivated equine rhinopneumonia vaccine prepared of the rhinopneumonia viral strain CB/69, an inactivated strangles vaccine prepared of the bacterial strain Six. equi H-34. |
 Sialic acid for bolstering immune system in elderly age / 2506087Present invention generally refers to immunology, particularly to bolstering the immune system in the elderly age and represents a composition. The composition for treating and preventing the diseases related to the altered immune system specified in a group consisting of infections, particularly bacterial, viral, mycotic and/or parasitic infections; the autoimmune diseases; the inborn immunity diseases, e.g. NK cell deficiency, phagocyte deficiency, such as leukocyte adhesion deficiency, cyclic neutropenia; and combinations thereof containing a protein fraction comprising acetylneuraminic acid related to a threonine-rich peptide/protein frame. |
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Method for preparing cell-free vaccine for pertussis immunisation / 2504399 Invention refers to medicine, namely to immunology and may be used for preparing an cell-free vaccine for pertussis immunisation. For this purpose, the strains Bordetella pertussis are grown on casein-carbon agar with microbial cell washout from the surface of a nutrient medium; the microbial cell concentration in the prepared suspension is reduced to 60-80 IU and pH 8.4±0.1. The complex of Bordetella pertussis antigens is extracted in 0.5% sodium deoxycholate to be reduced from the microbial cells by centrifugation. That is followed by diafiltration purification from the low-molecular fertiliser additives. The complex of Bordetella pertussis antigens is transferred into succinate-borate buffer solution 0.005M having pH 7.3±0.2 containing 0.09 M NaCl. The additional purification is ensured by column chromatography with anion-exchange resin Whatman DE-52 balanced by 0.005M succinate-borate buffer solution with pH 7.3±0.2 containing 0.09 M NaCl. The purified complex of Bordetella pertussis antigens is deactivated by 0.100±0.025% formalin at temperature (33±2)°C for 12±2 days. |
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Pharmaceutical composition containing enzyme ribonuclease and glycyrrhizic acid or salts thereof: ammonium or dipotassium or trisodium glycyrrhizinate / 2504396 Invention refers to pharmaceutical industry and represents a composition for treating wounds, ulcers, erosions, burns, freeze burns adhesions, as well as infections caused by the gram-positive and gram-negative bacteria Staphylococcus spp., Streptococcus spp., Enterococcus spp., Shigella spp., Escherichia spp., Salmonella spp., Proteus spp., Acinetobacter spp., Citrobacter spp., Pseudomonas spp., Serratia spp., Klebsiella spp., Antracoides spp., Cryptococcus spp., pathogenic fungi of the genera Microsporum, Trichophyton, Nocardia, Aspergillus, yeast-like fungi of the genus Candida (including the multiresistant strains), as well as Actinomycetes and some pathogenic protozoa (Entamoeba histolytica, Trichomonas vaginalis), containing an active ingredient in the form of 0.001-5.0 wt % of collagenase, 0.001-5.0 wt % of lysozyme and/or 0.001-5.0 wt % of sangvitirine, and an carrier in the form of 0.05-1.0 wt % of p-cyclodextrine or liposomes, and acceptable excipients. |
 Combined antibacterial preparation for treating acute intestinal infections / 2503451Invention refers to an antibacterial preparation for treating gastrointestinal diseases, preferentially acute intestinal infections, including unexplained. The declared preparation represents a mixture of an antibiotic and a biological ingredient, wherein the antibiotic is presented by chloramphenicol, while the biological ingredient is a complex of immunoglobulins IgG(56-60):IgA(16-22):IgM(22-24) in the following ratio, mg/capsule: chloramphenicol - 300 mg, the complex of immunoglobulins IgG,IgA,IgM - 120 mg. |
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Combined antibacterial preparation for treating acute intestinal infections / 2502509 Invention refers to an antibacterial preparation for treating gastrointestinal diseases, preferentially acute intestinal infections, including unexplained. The declared preparation represents a mixture of an antibiotic and a biological ingredient, wherein the antibiotic is presented by ofloxacin, while the biological ingredient is a complex of immunoglobulins IgG(56-60):IgA(16-22):IgM(22-24) in the following ratio, mg/flask: ofloxacin - 250, the complex of immunoglobulins IgG,IgA,IgM - 300. |
 Lactobacillus paracasei subspecies paracasei strain having antimicrobial and immunomodulating properties, and food product on its basis / 2501850Strain is deposited in CNCM and has number 1-3689. Strain has ability to inhibit the growth of pathogenic microorganisms in culture, and namely Escherichia coli, Salmonella enteritidis and Listeria monocytogenes. Strain has anti-inflammatory properties at the ratio of IL-10/IL-12 equal to 11.8. |
 Oxazolidinone derivatives / 2501799Invention relates to compounds of formula (I) where "----" denotes a bond or is absent; R1 is a C1-4alkoxy group or halogen; R1b is H or C1-3alkyl; U and V each independently denote CH or N; W is CH or N, or, if "----" is absent, W is CH2 or NH; under the condition that at least one of U, V and W is CH or CH2; A is -CH2-CH(R2)-B-NH-* or -CH(R3)-CH2-N(R4)-[CH2]m-*; where asterisks indicate a bond which binds said fragments through a CH2-group with an oxazolidinone fragment; B is CH2 or CO; and R2 is hydrogen, OH or NH2; R3 and R4 both denote hydrogen, or R3 and R4 together form a methylene bridge; m equals 0, 1 or 2; and G is a phenyl which is monosubstituted in position 3 or 4, or disubstituted in positions 3 and 4, where each substitute is independently selected from a group comprising C1-4alkyl, C1-3alkoxy group and halogen; or G is a group selected from groups G1 and G5 where M is CH or N; Q' is S or O; Z1 is N, Z2 is CH and Z3 is CH; or Z1 is CH, Z2 is N and Z3 is CH or N; or Z1 is CH, Z2 is CR5 and Z3 is CH; or Z1 is CH, Z2 is CH and Z3 is N; and R5 is hydrogen or fluorine; or a pharmaceutically acceptable salt thereof. The compound of formula (I) or a pharmaceutically acceptable salt thereof are used as a medicinal agent for preventing or treating bacterial infections. |
 Using pentaaminofullerenes as antimicrobial agents and antimicrobial composition based thereon / 2501785In formula 1 , X denotes a negative charge which is localised on a fullerene skeleton, a chlorine atom bonded to a carbon skeleton or a hydrogen atom; the NR1R2 moiety denotes an amine residue, where R1 and R2 are hydrogen atoms or linear or branched alkyl radicals (CmH2m+1; n=1-20) that are substituted with protonated (NH3 +) or unprotonated (NH2) amine groups, or a piperazine residue of general formula 1c-1 , where R, R'1, R'2, R'3 and R'4 are hydrogen atoms or linear or branched alkyl (CmH2m+1; n=1-20) radicals, as well as residues of aliphatic alcohols -(CH2)nOH, ethers -(CH2)nOR'5, thiols -(CH2)nSH, acids -(CH2)nCOOH, esters thereof -(CH2)nCOOR'5 or amides -(CH2)nCONR'5R'6, for which n=0-20, R'5 and R'6 are hydrogen atoms or linear alkyl (CmH2m+1; n=1-20) radicals. |
 Antifungal crystalline compounds / 2500679Described are novel crystalline forms of (1R,2R)-7-chloro-3-[2-(2,4-difluorophenyl)-2-hydroxy-1-methyl-3-(1H-1,2,4-triazol-1-yl)propyl]quinazolin-4(3H)-one: Form I, Form II, Form III, Form IV and Form VI, each characterised by X-ray powder diffraction (XRPD) data and infrared spectrum data, and a method of producing crystalline Form VI. The preferred form is Form VI, which has antifungal and antimicrobial activity, has the least impurity content, the highest uniform quality of the product, the highest uniform physical characteristics, including colour, rate of dissolution, easiness of handling and longest stability compared to the amorphous form. |
 Method of virus treatment by ultracentrifugation in sugar concentration gradient (versions) / 2503719Treatment methods of inactivated virus or fragmented virus (versions) are proposed. Methods involve addition of virus preparation to sugar gradient and further centrifugation to obtain peak fraction. Then, peak fraction is extracted to obtain cleaned virus. With that, sugar gradient is obtained by continuous ultracentrifugation of the first and the second buffer sugar solution. Besides, the first buffer sugar solution includes the first physiological buffer, and the second buffer sugar solution includes the second physiological buffer that is the same or different from the first physiological buffer. As per one version, sugar concentration in the first buffer solution is equivalent to saccharose concentration of 35 to 50 wt %, and sugar concentration in the second buffer solution is equivalent to saccharose concentration of 50 to 65 wt %. Besides, the second buffer sugar solution has higher density in comparison to that of the first one. As per the other method version, density of the first buffer sugar solution is 1.15 kg/l to 1.23 kg/l, and density of the second one is 1.23 kg/l to 1.32 kg/l. |
FIELD: biotechnologies.
SUBSTANCE: vaccine strain for influenza virus A/17/mallard/Netherlands/00/95(H7N3) is proposed. Strain A/17/mallard/Netherlands/00/95(H7N3) is harmless for laboratory animals. Strain A/17/mallard/Netherlands/00/95(H7M3) has antigenic actuality and attenuation thus meeting the requirements to vaccine strains included in influenza live vaccine, and may be used for production of influenza live and inactivated intranasal vaccine in health care service for influenza prevention.
EFFECT: temperature sensitivity and high degree of cold adaptability.
7 tbl, 3 ex
The invention relates to medical Virology and can be used in public health for prevention of influenza caused dangerous viruses of avian influenza. For the production of intranasal live influenza vaccine (LFV) or inactivated influenza vaccine (EBP) proposed influenza virus strain A/17/mallard/Netherlands/00/95(H7N3).
Despite the wide spread of the pandemic influenza virus subtype H1N1, avian influenza viruses of subtypes A(H5N1), A(H7N1) and A(H9N2) continue to cause outbreaks of disease among the people. Viruses of subtype H7 can be widely disseminated among poultry, and acquiring properties increased pathogenicity as a result of additional adaptation and recombination process, as was shown during outbreaks caused by the A(H7N7), A(H7N1) and A(H7N4) in Europe and Australia. Need to develop appropriate vaccine preparations, including, and to fight with avian influenza viruses, reflected in the order of the RF Ministry of health [Order №40 MOH RF dated 28.12.2004].
Currently, the vaccine strains for the LFV is obtained by modern epidemic viruses with donor strains, resulting in having mixed genome. Donor And/Leningrad/134/17/57(H2N2) - strain of the influenza virus permitted to obtain harmless nasal vaccine for adults and children [Aleksandrova GI New in epidemiology and prevention of viral infections. HP, 1986. - P.66-83]. The genes encoding the haemagglutinin () and neiraminidazu (NA), inherited from actual epidemic strain, and six genes internal and nonstructural proteins (2, 1, PA, NP, M, NS) from harmless HA donor . From the received reassortants choose vaccine candidate with the formula genome 6:2 that will meet the requirements of the antigenic specificity, characteristic for the parent virus «wild» type, as well as the respective characteristics of cold adaptation, reproduction ability at low temperature.
Currently used for prevention of the epidemic of influenza And vaccine strains of H1N1 and H3N2 subtypes cannot call a defensive reaction in case of large-scale outbreaks caused by influenza viruses of subtype hemagglutinin subtype H7, to which the vast majority of the population has no immunity.
Currently known strain A/17/duck/Potsdam/86/92(H5N2) [Patent no 2318871, publ. 10.03.2008], intended for the production of live and inactivated vaccines against avian influenza subtype H5. But he doesn't have a preventive effect in case of spread of the influenza virus subtype H7.
Abroad for the preparation of the vaccine strain LFV subtype H7-based donor strain A/Ann arbor/6/60(H2N2) was used highly pathogenic virus A/Netherlands/219/03(H7N7) [Min J., J. Virol. 2010 Nov; 84(22): 11950-60].
Also known strain A/17/wild duck/Netherlands/00/84(H7N3)obtained by the methods of classical genetic in developing chicken embryos () on the basis of donor strain Linen/17 [Cheap A., - 2009. - №1. - P.31-36]. However, this strain is not quite satisfy the requirements for vaccine strains included in the composition of the LFV for immunization of people. First of all, the strain was not enough : the difference in reproductive activity and reduced down to 25 degrees temperature for the strain of A/17/wild duck/Netherlands/00/84(H7N3) was 4.2 Ig EID 50 (rct 25-sign), while it is established that for vaccine strains, this difference should not exceed 3.0 Ig EID 50 [Science, 1959. - Vol.129. No. 3358. - .1287-1288]. In addition, direct sequencing shows the presence of the nucleotide replacement G > A in position 543 gene ON strain A/17/wild duck/Netherlands/00/84(H7N3) compared with the gene to THE «wild» virus of avian influenza. The specified replacement was accompanied by amino acid substitution of glycine on acid in position 214, that led to a change of polarity and charge amino acids near plot. Such replacements are undesirable for vaccine strains, as they can negatively affect their antigenic specificity, causing a decrease in the protective efficacy of vaccination.
Since the task to be solved by the claimed invention is to obtain a fully vaccine strain with subtype H7 for immunization of people were held for more stages of cloning. This method was chosen so called «short passages» [Fazekas de St. Groth, Intervirology, 1975. - Vol.5. - P.335-341] in cold temperatures, when the mixture viral clones in a sensitive system (chicken embryos, EC) in the presence of factors in shortened period of incubation. A mixture of viral clones received after crossing the donor And/Leningrad/134/17/57(H2N2) and virus of the bird influenza A/mallard/Netherlands/12/00 (H7N3) in cavity 10-11-day-old chick embryos (EC), the past 2 selective passage and one cloning in the presence of antiserum to the donor And/Leningrad/134/17/57(H2N2) and reduced down to 25 degrees incubation temperature, optional cloned 2 times reduced to 30oC temperature for a shortened up to 28 hours period, increasing the final dilution up to 1:10 9 . With the help of selection of these conditions could not allocate a clone A/17/mallard/Netherlands/00/95(H7N3), which differed in their characteristics from the others, such as most pronounced signs of cold adaptation, because the level of its reproduction reduced down to 25 degrees temperature was 100 times higher than that obtained earlier viruses (table 1).
Table 1Reproduction in low temperature clones A(H7N3)
(№ clone)
The number of passages in the process of
Reproductive activity EID 50 /0.2 ml
RCT 25 EID 50 /0.2 ml
34° 34° 8/4 68,1 approximately 0.5
4,1±0,4 4,0 8/6 6 8,7±0,2 4,7±0,2 4,0 8/10 6 8,3±0,1 4,0±0,1 4,39/5 AL 7/mallard/Netherlands/00/95(H7N3)
7 (including 2 «shortened»)
8,7±0,2 6,4±0,2 2,3High level of cold adaptation reassortant And/17/mallard/Netherlands/00/95(H7N3) was based on molecular-genetic methods. Sequencing strain revealed the presence in the gene internal protein NP additional nucleotide substitution With a 1066 -, which resulted in amino acid replacement Leu-341-lie. This replacement was previously detected in advanced attenuated donor strain, And/Leningrad/134/47/57(H2N2), which was obtained after a number of passages strain A/Leningrad/134/17/57(H2N2) in FE at low temperature and later was widely used for a number of years in preparing the LFV for immunization of young children [Klimov, et al., 1992].
Analysis of the cloned vaccine strain A/17/mallard/Netherlands/00/95(H7N3) has been implemented using the restriction analysis of DNA copies of RNA segments, obtained by back-polymerase chain reaction (RT-PCR) [A.I. Klimov, J. Virol. Method. - 1995. - №55. - P. 445-446]. Obtained by RT-PCR DNA copy of the segments 2, 1, PA, NP, M and NS were treated accordingly processed by endonucle ases restriction Tru9l, Hindlll, BamHI, Asnl, EcoRI, Pvul, Ncil, respectively. Additional copies segments 1, PA and M were treated restrictases BstXI, Asnl and BciVI. Restriction analysis of DNA copies RNA segments (Table 2) and direct sequencing showed that And/17/mallard/Netherlands/00/95 (H7N3) has the composition of the genome 6:2. All coding nucleotide substitution, characteristic for the donor A/Leningrad/134/17/57(H2N2), found in the genes of internal and nonstructural proteins strain A/17 a/mallard/Netherlands/00/95 (H7N3).
sequencing of the gene ON virus A/17/mallard/Netherlands/00/95(H7N3) revealed no differences in structure from the parent virus «wild» type. Spent sequencing of the gene NA virus A/17/mallard/Netherlands/00/95(H7N3) also did not reveal differences in structure from the parent virus «wild» type.
Thus, the received strain is characterized by a combination required for vaccine strains LFV symptoms: cold adaptation and identity of surface antigens parent virus to wild type. At the same time, the resulting strain possesses high in chicken embryos (>10 9 EID 50 /ml), that allows to use it in production of inactivated influenza vaccine (EBP), requires that you obtain a large number of viral material for its further purification and concentration.
Affiliation hemagglutinin parent strain of «wild» type A/mallard/Netherlands/12/00(H7N3) reassortant was confirmed in reaction haemagglutination inhibition (). The study of the antigenic properties shows that avian influenza virus A/mallard/Netherlands/12/00 (H7N3) and it HA are identical (interaction with homologous serum according to TU 42-14-79-79). Indicators of interaction reassortant And/17/mallard/Netherlands/00/95(H7N3), as well as parental strain of the A/mallard/Netherlands/12/00(H7N3) with antisera ferrets obtained to viruses, selected from people during the outbreak in the Netherlands in 2003, including the case of the lethal infection (And/Netherlands/219/03), differed from homologous titles corresponding antigens not more than 2 times (table 2).
Table 3Nucleotide substitution found in the genes of internal and nonstructural proteins strain A/17/mallard/Netherlands/00/95 (H7N3) before and after passage in chicken embryos
VirusCharacteristic
Nucleotide positions in the internal genes
2 1 PA m NS 1459 819 1795 107 1045 68 969 798A/Leningrad/134/57 (H2N2)
Epidemic virus
G G G T G And G GA/Leningrad/134/17/57 (H2N2)
HA donor
T T And With T G And AndA/17 a/mallard/ Netherlands/00/95 (H7N3) to passages in the EC
vaccinal strain
T T And C T G And AndA/17/mallard/Netherlands/00/95(H7N3) after 5 passages in TBE
T T And C T G And AndTable 4 presents the indicators of reproduction in the EC strain A/17/mallard/Netherlands/00/95(H7N3). It is shown that And/17/mallard/Netherlands/00/95(H7N3) showed high reproductive activity at the optimal temperature of incubation 34°N Like HA parent strain And/Leningrad/134/17/57(H2N2), strain A/17 a/mallard/Netherlands/00/95(H7N3) well reduced to 25-26°With temperature and almost lost the ability to reproduce at 40 C (ts-ca - phenotype). The parent virus «wild» type A/mallard/Netherlands/12/00(H7N3), on the contrary, different temperature resistance and the lack of reproduction at 25 deg C.
Table 4Reproductive activity reassortant And/17/mallard/Netherlands/00/95(H7N3) at different temperatures incubation in chicken embryos
Virus (feature)
Reproductive activity at a temperature (Ig EID 50 /1 ml)
Phenotype
34° 40 C 25 OC 26 CA/17/mallard/Netherlands/00/95(H7N3) (6:2 )
9,2 approximately 0.5
1,8±0,26,9 approximately 0.5
7,5±0,1 Ts, caA/Leningrad/134/17/57(H2N2) (HA donor )
9,5±0,3of 1.6±0.1
7,0±0,1 8,7±0,6 7s, caThe a/mallard/Netherlands/12/00 (H7N3) ( avian strain)
8,7±0,2 8,0±0,2 <1,5of 1.6±0.1
Non-ts, non-ca
Therefore, the vaccine strain A/17/mallard/Netherlands/00/95(H7N3) is characterized by a combination of useful features necessary strain - antigenic specificity of the virus hemagglutinin «wild» type A/mallard/Netherlands/12/00(H7N3), structure of the genome, necessary for vaccine strains , a good adaptation to low temperature cultivation, which correlates with characteristic of the attenuated donor strain. Characteristics of the new strain- and high reproductive activity in chicken embryos - allow its use for the production of live and inactivated influenza vaccine.
Strain deposited in the collection of the Scientific research Institute of Virology them. .. Ivanovo RAMS # 2717. Morphology of strain - polymorphic, typical for influenza virus.
CHARACTERISTICS OF THE RESULTING STRAIN.
The infectious activity of reproduction in developing chicken embryos, with a 33 degree for 48 hours at 9.2 Ig EID 50 / 0,2 ml
activity - 1:256.
Passport of the vaccinal strain A/17/mallard/Netherlands/00/95(H7N3)
attached (p.7).
Results of pre-clinical study of the proposed strain.
Example 1. Study of the safety of the reassortant And/17/mallard/Netherlands/00/95 (H7N3) carried out the hens white Leghorn breed. Each of the eight 5-week-old chickens injected intravenously 0.2 ml liquid with infectious activity of the virus A/17/mallard/Netherlands/00/95(H7N3) 10 8,1 EID 50 /0,2 ml within 10 days conducted monitoring for signs of infection (respiratory infection, weakness, diarrhea, cyanosis in the field of inoculation, swelling of the head, neurological symptoms). Index of pathogenicity counted as 0 - no symptoms, 1 - development of the signs of infection, 2 - severe infection, 3 - a lethal outcome.
Within 10 days after the on/in the inoculation strain A/17/mallard/Netherlands/00/95(H7N3) no animal has not been registered signs of infection (pathogenicity index 0). Thus, reassortant/17/mallard/Netherlands/00/95(H7N3) was harmless intravenous chickens.
On the third day after intranasal dose of 6 Ig EID 50 /ml And/17/mallard/Netherlands/00/95(H7N3) has not been isolated from and the procedure.
Virus isolation And/17/mallard/Netherlands/00/95(H7N3) of the procedure after intranasal chickens
no animal
Cloaca 1001≤10 0.9 /ml
≤10 0.9 /ml
1002≤10 0.9 /ml
≤10 0.9 /ml
1003≤10 0.9 /ml
≤10 0.9 /ml
1004≤10 0.9 /ml
≤10 0.9 /ml
1005≤10 0.9 /ml
≤10 0.9 /ml
1006≤10 0.9 /ml
≤10 0.9 /ml
1007≤10 0.9 /ml
≤10 0.9 /ml
1008≤10 0.9 /ml
≤10 0.9 /ml
1009≤10 0.9 /ml
≤10 0.9 /ml
1010≤10 0.9 /ml
≤10 0.9 /ml
Within 14 days was not observed for signs of infection, the virus was not isolated from and swabs and tissues of the lung, kidney, heart and brain. On the 14th day after intranasal not registered . Thus, the effect of the introduction of virus A/17/mallard/Netherlands/00/95(H7N3) chickens was similar to the action of the seasonal influenza viruses with a low risk of infection.
Example 2. The proposed vaccine strain A/17/mallard/Netherlands/00/95 (H7N3) harmless for mice and Guinea pigs.
Histological study shows that the introduction of the vaccine strain A/17 a/mallard/Netherlands/00/95 laboratory animals - mice, not causes a defensive reaction in the form of inflammation, alterative manifestations in the form of dystrophic and destructive lesions in the liver, kidney, spleen, heart muscle - myocardium, stromalnom component of the internal organs. In addition, it is not marked dystrophic changes in neurons, glial elements and circulatory disorders of microcirculation of the brain. Thus, the use of the vaccine does not cause morphological changes, which indicates a good tolerability and safety of the vaccine And/17/mallard/Netherlands/00/95.
Example 3. Vaccinal strain A/17 a/mallard/Netherlands/00/95(H7N3) for mice.
It is shown that the vaccine strain A/17/mallard/Netherlands/00/95(H7N3) is immunogenic properties by intra mice. To compound titles antibodies increase in the group of immunized mice, the two-time immunization is more effective.
Results with serum mice immunized intranasal 10 7 Ig EID 50 strain A/17/mallard/Netherlands/00/95(H7N3) 4 weeks after immunization
Group GTSTo immunization
Once Twice LFV 5,0 8,7 40,0Placebo (solution FB)
5,0 5,0 5,0Influenza virus strain A/17/mallard/Netherlands/00/95(H7N3), in the State collection of viruses Scientific research Institute of Virology them. .. Ivanovo RAMS # 2717, for the production of live intranasal and production of inactivated influenza virus vaccines.