ABSTRACT
The WHO approved protocol of pandemic influenza virus A/H1N1 identification by the method of one-step RT-PCR was adopted. The cost of research was decreased due to adoption of RNA purification method and utilizing of the common enzyme systems for RT-PCR.
Subject(s)
Disease Outbreaks , Influenza A Virus, H1N1 Subtype/isolation & purification , Influenza, Human/virology , Reverse Transcriptase Polymerase Chain Reaction/methods , Diagnosis, Differential , Humans , Influenza A Virus, H1N1 Subtype/genetics , Influenza, Human/epidemiology , Mouth Mucosa/virology , Sensitivity and Specificity , UkraineABSTRACT
A diagnostic kit for detection of avian influenza virus by real-time polymerase chain reaction was developed. This kit allows to identify the influenza viruses type A and highly pathogenic strain of avian influenza virus H5N1. The diagnostic kit is universal and adopted for ABI PRISM SDS (Applied Biosystems), RotorGene (Corbett Research) and iQCycler (BioRad) PCR machines.
Subject(s)
Influenza A Virus, H5N1 Subtype , Influenza in Birds/virology , Polymerase Chain Reaction/methods , Amino Acid Sequence , Animals , Birds , Influenza A Virus, H5N1 Subtype/genetics , Influenza A Virus, H5N1 Subtype/isolation & purification , Influenza A Virus, H5N1 Subtype/pathogenicity , Molecular Sequence Data , RNA, Viral/genetics , Reproducibility of Results , Sensitivity and Specificity , VirulenceABSTRACT
The chemical synthesis of a modified gene fragment that codes IgG-binding domain of protein G of Streptococcus sp. (SPG) was done. Two copies of gene fragmentes SPG were cloned into plasmid vector pET-24(a), for expression of recombinant truncated protein G (rSPG) in the culture of Escherichia coli. IgG-binding capacity and specificity for rSPG was confirmed by model assay such interaction in vitro. Basing on the obtained data we affirm that truncated recombinant protein G is able to bind immunoglobulin G in mammalians and can be used for purification of IgG from sera by affinity chromatography or for synthesis of immunoassay conjugates with broad range specificity.
Subject(s)
Bacterial Proteins , Immunoenzyme Techniques/methods , Immunoglobulin G/metabolism , Recombinant Proteins , Streptococcus/metabolism , Bacterial Proteins/biosynthesis , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Cloning, Molecular , Electrophoresis, Polyacrylamide Gel , Escherichia coli/genetics , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Streptococcus/geneticsABSTRACT
Several Escherichia coli strains producing HIV-specific recombinant proteins including env-1, env-2, and gag sequences have been obtained using different gene engineering techniques. These proteins have been isolated and purified from bacterial lysates by ion exchange chromatography on DEAE-Toyopearl and Streamline-SP columns. Polyacrylamide gel electrophoresis and immunoblotting have been used to prove recombinant proteins purity and to identify them. These proteins have been shown to be adequate as antigen preparations for enzyme immunoassay test-systems aimed at anti-HIV antibodies detection.
Subject(s)
HIV-1/metabolism , HIV-2/metabolism , Viral Proteins/biosynthesis , Chromatography, DEAE-Cellulose , Electrophoresis, Polyacrylamide Gel , Escherichia coli/metabolism , Genetic Engineering/methods , Immunoblotting , Immunochemistry , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Recombinant Proteins/isolation & purification , Viral Proteins/chemistry , Viral Proteins/isolation & purificationABSTRACT
The results from comparative studies in the reactions of immunofluorescence, complement binding, diffusion precipitation, hemagglutination, solid-phase immunoenzyme analysis, histochemical variant of immunoenzyme analysis as tests for detection of cattle rotavirus in the process of its isolation from pathological material and adaptation to cell cultures are presented. The immunofluorescence reaction is shown to have an advantage over the other reactions.
