Investigation of the causative agents of the 1982 Gazankulu poliomyelitis outbreak, using four biochemical techniques
- Authors: Gibson, Katherine Margaret
- Date: 1989
- Subjects: Poliomyelitis -- Analysis , Poliomyelitis -- History -- South Africa , Poliomyelitis vaccine -- Analysis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3878 , http://hdl.handle.net/10962/d1001612
- Description: Comparison of poliovirus strains was carried out to determine the origin of the virus in two isolates obtained during the 1982 outbreak of poliomyelitis in Gazankulu. Comparisons of the outbreak isolates with vaccine and wild-type strains of the same poliovirus type were carried out using four biochemical techniques. SDS-polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional thin-layer chromatography (TLC) and reversed-phase high-performance liquid-chromatography (RP-HPLC) were used for comparing viral capsid proteins. Comparison of poliovirus strains at a genetic level was carried out using two-dimensional oligonucleotide mapping of viral RNA. Results showed the type 1 poliovirus isolate, 5061, to be a novel wild-type poliovirus. The type 2 isolate, 5068, was closely related to the poliovirus type 2 Sabin vaccine strain, P712. It was concluded that the intrinsic variability of poliovirus strains was responsible for the appearance of isolate 5068
- Full Text:
- Date Issued: 1989
- Authors: Gibson, Katherine Margaret
- Date: 1989
- Subjects: Poliomyelitis -- Analysis , Poliomyelitis -- History -- South Africa , Poliomyelitis vaccine -- Analysis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3878 , http://hdl.handle.net/10962/d1001612
- Description: Comparison of poliovirus strains was carried out to determine the origin of the virus in two isolates obtained during the 1982 outbreak of poliomyelitis in Gazankulu. Comparisons of the outbreak isolates with vaccine and wild-type strains of the same poliovirus type were carried out using four biochemical techniques. SDS-polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional thin-layer chromatography (TLC) and reversed-phase high-performance liquid-chromatography (RP-HPLC) were used for comparing viral capsid proteins. Comparison of poliovirus strains at a genetic level was carried out using two-dimensional oligonucleotide mapping of viral RNA. Results showed the type 1 poliovirus isolate, 5061, to be a novel wild-type poliovirus. The type 2 isolate, 5068, was closely related to the poliovirus type 2 Sabin vaccine strain, P712. It was concluded that the intrinsic variability of poliovirus strains was responsible for the appearance of isolate 5068
- Full Text:
- Date Issued: 1989
A novel adjuvant : polymerised serum albumin beads
- Authors: Dewar, John Barr
- Date: 1985
- Subjects: Antigens , Serum albumin
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4104 , http://hdl.handle.net/10962/d1011146 , Antigens , Serum albumin
- Description: Lee, T. et al (1981) proposed the encapsulation of hormones such as progesterone into serum albumin beads, such that their in vivo proteolysis would allow a gradual release of hormone at low levels, for extended hormone action. It was proposed, in the Department of Microbiology, Rhodes University, to replace the hormone component of the above bead formulation, with virus as antigen, in the development of a vaccine. Beads optimally crosslinked at 1% final glutaraldehyde concentration, containing Nodamura virus, were shown to promote an adjuvant effect in vivo, analogous to the release of antigen from Freund's Complete Adjuvant (FCA), so that extended immunostimulation resulted. It was shown that soluble antigen promoted a short-lived primary immune response, peaking around day 25 following inoculation. Antigen presented in beads, on the other hand, initially elicited a low humoral response, but this response gradually increased up to a peak around day 110 post inoculation, before decreasing. No apparent adverse side-effects were noted following inoculation of antigen-containing serum albumin beads, compared to necrosis following antigen in FCA inoculation, supporting the proposal of using albumin homotypic for the test inoculee animal, so that the beads would themselves be non-immunogenic and would merely act as a vehicle in the vaccine formulation. The indirect enzyme-linked immunosorbent assay (ELISA) was used to monitor the humoral response to antigen following inoculation. Results showed that covalent crosslinking of albumin in the formation of the beads did not promote immunogenicity on the part of the chemically altered albumin. The ELISA test was used to indicate the kinetics of the IgG response to Nodamura virus when presented in formulations such as: Freely soluble virus or its subunit; soluble intact virus inactivated by treatment with glutaraldehyde; intact virus entrapped in serum albumin beads cross; linked at different percentage final glutaraldehyde concentrations and also virus subunit prepared in albumin beads. The presence of virus-neutral ising antibodies was noted in serum obtained from rabbits inoculated with virus entrapped in albumin beads. Virus infectivity, titrated in mice, showed protection against virus challenge after incubation of virus with serum obtained above.
- Full Text:
- Date Issued: 1985
- Authors: Dewar, John Barr
- Date: 1985
- Subjects: Antigens , Serum albumin
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4104 , http://hdl.handle.net/10962/d1011146 , Antigens , Serum albumin
- Description: Lee, T. et al (1981) proposed the encapsulation of hormones such as progesterone into serum albumin beads, such that their in vivo proteolysis would allow a gradual release of hormone at low levels, for extended hormone action. It was proposed, in the Department of Microbiology, Rhodes University, to replace the hormone component of the above bead formulation, with virus as antigen, in the development of a vaccine. Beads optimally crosslinked at 1% final glutaraldehyde concentration, containing Nodamura virus, were shown to promote an adjuvant effect in vivo, analogous to the release of antigen from Freund's Complete Adjuvant (FCA), so that extended immunostimulation resulted. It was shown that soluble antigen promoted a short-lived primary immune response, peaking around day 25 following inoculation. Antigen presented in beads, on the other hand, initially elicited a low humoral response, but this response gradually increased up to a peak around day 110 post inoculation, before decreasing. No apparent adverse side-effects were noted following inoculation of antigen-containing serum albumin beads, compared to necrosis following antigen in FCA inoculation, supporting the proposal of using albumin homotypic for the test inoculee animal, so that the beads would themselves be non-immunogenic and would merely act as a vehicle in the vaccine formulation. The indirect enzyme-linked immunosorbent assay (ELISA) was used to monitor the humoral response to antigen following inoculation. Results showed that covalent crosslinking of albumin in the formation of the beads did not promote immunogenicity on the part of the chemically altered albumin. The ELISA test was used to indicate the kinetics of the IgG response to Nodamura virus when presented in formulations such as: Freely soluble virus or its subunit; soluble intact virus inactivated by treatment with glutaraldehyde; intact virus entrapped in serum albumin beads cross; linked at different percentage final glutaraldehyde concentrations and also virus subunit prepared in albumin beads. The presence of virus-neutral ising antibodies was noted in serum obtained from rabbits inoculated with virus entrapped in albumin beads. Virus infectivity, titrated in mice, showed protection against virus challenge after incubation of virus with serum obtained above.
- Full Text:
- Date Issued: 1985
A study of the molecular variation between orbivirus proteins
- Authors: Whistler, Toni
- Date: 1985 , 2013-03-13
- Subjects: Proteins -- Analysis , Polypeptides , Bluetongue virus , Orbivirus infections
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3892 , http://hdl.handle.net/10962/d1003290 , Proteins -- Analysis , Polypeptides , Bluetongue virus , Orbivirus infections
- Description: The aim of this study was to initiate a structural analysis of the capsid polypeptides from several serotypes of bluetongue virus in order to provide insight into the relatedness and possible origins of the different serotypes. Tryptic peptide mapping of ¹²⁵I-labelled group antigen by ion exchange chromatography was used to assess the structural relatedness of seven BTV serotypes from Southern Africa, North America and Australia. Each serotype had several tyrosine containing tryptic peptides which were unique, but approximately 35% of the peptides analyzed were found to be highly conserved between all 7 serotypes. BTV-20 appeared to be closely related to BTV-B and these two serotypes with BTV-4 and BTV-17 appeared to form a closely knit central cluster. , KMBT_363 , Adobe Acrobat 9.53 Paper Capture Plug-in
- Full Text:
- Date Issued: 1985
- Authors: Whistler, Toni
- Date: 1985 , 2013-03-13
- Subjects: Proteins -- Analysis , Polypeptides , Bluetongue virus , Orbivirus infections
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3892 , http://hdl.handle.net/10962/d1003290 , Proteins -- Analysis , Polypeptides , Bluetongue virus , Orbivirus infections
- Description: The aim of this study was to initiate a structural analysis of the capsid polypeptides from several serotypes of bluetongue virus in order to provide insight into the relatedness and possible origins of the different serotypes. Tryptic peptide mapping of ¹²⁵I-labelled group antigen by ion exchange chromatography was used to assess the structural relatedness of seven BTV serotypes from Southern Africa, North America and Australia. Each serotype had several tyrosine containing tryptic peptides which were unique, but approximately 35% of the peptides analyzed were found to be highly conserved between all 7 serotypes. BTV-20 appeared to be closely related to BTV-B and these two serotypes with BTV-4 and BTV-17 appeared to form a closely knit central cluster. , KMBT_363 , Adobe Acrobat 9.53 Paper Capture Plug-in
- Full Text:
- Date Issued: 1985
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