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Molecular characterization of Mycoplasma synoviae in chickens in South Africa using single-stranded conformation polymorphism and high-resolution melting curve analysis of the vlhA gene
Dissertation (MSc)--University of Pretoria, 2012.
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| Format: | Thesis |
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University of Pretoria
2013
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| _version_ | 1867613725145432064 |
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| access_status_str | Open Access |
| author2 | Venter, Estelle Hildegard |
| author_browse | Venter, Estelle Hildegard |
| author_facet | Venter, Estelle Hildegard |
| collection | Thesis |
| dc_rights_str_mv | © 2012 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria |
| description | Dissertation (MSc)--University of Pretoria, 2012. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/26196 |
| institution | University of Pretoria (South Africa) |
| last_indexed | 2026-06-10T12:40:42.021Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2013 |
| publishDateRange | 2013 |
| publishDateSort | 2013 |
| publisher | University of Pretoria |
| publisherStr | University of Pretoria |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/26196 Molecular characterization of Mycoplasma synoviae in chickens in South Africa using single-stranded conformation polymorphism and high-resolution melting curve analysis of the vlhA gene Venter, Estelle Hildegard Bisschop, Shahn P.R. mashikoane@webmail.co.za Raphela, Mashikoane Pinky Jane Chickens South africa Vlha gene Mycoplasma synoviae UCTD Dissertation (MSc)--University of Pretoria, 2012. Mycoplasma synoviae (Ms) causes respiratory infection and synovitis in chickens and turkeys and is an economically important pathogen of poultry worldwide. It is critically important to characterize Ms strains, especially in countries in which poultry flocks are vaccinated with the live attenuated Ms strain MS-H. Vaccination with this vaccine may cause sero-conversion and persistence of the vaccine strain in the respiratory tract and will frequently result in positive Ms cultures and PCR results. Vaccination of flocks therefore complicates the diagnosis of Ms by the presence of detectable antibodies in the blood. Many diagnostic techniques cannot distinguish between the vaccine strain and wild type strain. Single stranded conformation polymorphism (SSCP) and real-time PCR with high melting curve (HRM) analysis can discriminate between the different Ms strains obtained from the field and also distinguish them from the live vaccinestrains. These techniques provide effective tools for the further study of the epidemiology and spread of Ms strains in chickens in South Africa. This project was undertaken to establish whether SSCP and HRM analyses can be used effectively to discriminate between Ms field isolates and the vaccine strain. Mycoplasma synoviae DNA was extracted from samples and conventional PCR was performed using oligonucleotide primers complementary to the single-copy conserved 5’ end of the variable lipoprotein and haemagglutinin encoding gene (vlhA). Twenty six samples were separated by agarose gel electrophoresis and prepared for SSCP and real-time PCR and HRM curve analysis. Results obtained from SSCP were compared to real-time PCR/HRM. Differences obtained by SSCP and melting curve analysis between different isolates were confirmed by sequencing. Results obtained from the different techniques differentiated the strains from the vaccine strain (isolate Ms10), which had a different melting temperature to the others and a different band pattern on the SSCP gel. These results confirmed that HRM and SSCP methods can be used to detect and discriminate between Mycoplasma synoviae field isolates and the vaccine strain. Veterinary Tropical Diseases unrestricted 2013-09-07T03:43:40Z 2013-07-12 2013-09-07T03:43:40Z 2013-04-12 2012 2013-07-10 Dissertation Raphela, MPJ 2012, Molecular characterization of Mycoplasma synoviae in chickens in South Africa using single-stranded conformation polymorphism and high-resolution melting curve analysis of the vlhA gene, MSc dissertation, University of Pretoria, Pretoria, viewed yymmdd < http://hdl.handle.net/2263/26196 > E13/4/675/gm http://hdl.handle.net/2263/26196 http://upetd.up.ac.za/thesis/available/etd-07102013-154154/ © 2012 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria application/pdf University of Pretoria |
| spellingShingle | Chickens South africa Vlha gene Mycoplasma synoviae UCTD Molecular characterization of Mycoplasma synoviae in chickens in South Africa using single-stranded conformation polymorphism and high-resolution melting curve analysis of the vlhA gene |
| title | Molecular characterization of Mycoplasma synoviae in chickens in South Africa using single-stranded conformation polymorphism and high-resolution melting curve analysis of the vlhA gene |
| title_full | Molecular characterization of Mycoplasma synoviae in chickens in South Africa using single-stranded conformation polymorphism and high-resolution melting curve analysis of the vlhA gene |
| title_fullStr | Molecular characterization of Mycoplasma synoviae in chickens in South Africa using single-stranded conformation polymorphism and high-resolution melting curve analysis of the vlhA gene |
| title_full_unstemmed | Molecular characterization of Mycoplasma synoviae in chickens in South Africa using single-stranded conformation polymorphism and high-resolution melting curve analysis of the vlhA gene |
| title_short | Molecular characterization of Mycoplasma synoviae in chickens in South Africa using single-stranded conformation polymorphism and high-resolution melting curve analysis of the vlhA gene |
| title_sort | molecular characterization of mycoplasma synoviae in chickens in south africa using single stranded conformation polymorphism and high resolution melting curve analysis of the vlha gene |
| topic | Chickens South africa Vlha gene Mycoplasma synoviae UCTD |
| url | http://hdl.handle.net/2263/26196 http://upetd.up.ac.za/thesis/available/etd-07102013-154154/ |