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Development of PCR-based methods for detection of African lyssaviruses
Dissertation (MSc)--University of Pretoria, 2010.
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University of Pretoria
2013
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| _version_ | 1867613684022378496 |
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| access_status_str | Open Access |
| author2 | Weyer, Jacqueline |
| author_browse | Weyer, Jacqueline |
| author_facet | Weyer, Jacqueline |
| collection | Thesis |
| dc_rights_str_mv | © 2010, 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, 2010. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/28545 |
| institution | University of Pretoria (South Africa) |
| last_indexed | 2026-06-10T12:40:02.981Z |
| 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:2263/28545 Development of PCR-based methods for detection of African lyssaviruses Weyer, Jacqueline Nel, Louis Hendrik Markotter, Wanda s23121719@tuks.co.za Coertse, Jessica African lyssaviruses Polymerase chain reaction (PCR) Pcr-based methods UCTD Dissertation (MSc)--University of Pretoria, 2010. The etiological agent of rabies encephalitis belongs to the genus Lyssavirus in the Rhabdoviridae family. Lyssaviruses are negative sense, single stranded RNA viruses and cause an estimated 55 000 human deaths per year with 44% of these deaths occurring in Africa (WHO, 2005). With intense research effort and increased sequence information it is becoming evident that the Lyssavirus genus is much more diverse than initially thought and therefore diagnostic methods need to be modified accordingly. The African continent sustains a diverse variety of lyssaviruses, however, most countries in Africa do not have active surveillance or necessary diagnostic tools and therefore rabies-related lyssaviruses are underreported. Previous studies have indicated that real-time PCR has improved sensitivity and rapidity over conventional molecular diagnostic methods with the added advantage of allowing accurate estimations of viral load in a wide variety of samples. Several realtime PCR assays have been developed; however, none were specifically aimed at detection of lyssaviruses present on the African continent. This study was therefore aimed at evaluating certain molecular diagnostic methods for the detection of African lyssaviruses. Furthermore, the application of real-time PCR for various fields in lyssavirus research i.e. diagnostics, surveillance and pathogenicity studies were evaluated. This study revealed two different hemi-nested PCR assays capable of detecting representatives of African lyssaviruses. A real-time PCR was developed that was successful for the detection of African lyssaviruses. In addition, a quantitative assay and internal control was successfully employed for confirming ante-mortem human rabies diagnosis as well as post-mortem animal rabies diagnosis in formalin fixed brain material. As such the real-time PCR assay developed in this study could therefore be routinely used for ante-mortem diagnosis and as a confirmatory test for post-mortem diagnosis. The ability of this assay to detect and quantify all currently known African lyssaviruses not only offers improved surveillance capacity, but offers unique potential as a sensitive tool to track virus movement in pathogenicity studies. These aspects are important in our search for a better understanding of the complex epidemiological and viral characteristics of African lyssaviruses. Copyright Microbiology and Plant Pathology unrestricted 2013-09-07T13:42:29Z 2010-10-08 2013-09-07T13:42:29Z 2010-04-28 2010-10-08 2010-10-08 Dissertation Coertse, J 2010, Development of PCR-based methods for detection of African lyssaviruses, MSc dissertation, University of Pretoria, Pretoria, viewed yymmdd < http://hdl.handle.net/2263/28545 > E10/481/gm http://hdl.handle.net/2263/28545 http://upetd.up.ac.za/thesis/available/etd-10082010-143715/ © 2010, 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 | African lyssaviruses Polymerase chain reaction (PCR) Pcr-based methods UCTD Development of PCR-based methods for detection of African lyssaviruses |
| title | Development of PCR-based methods for detection of African lyssaviruses |
| title_full | Development of PCR-based methods for detection of African lyssaviruses |
| title_fullStr | Development of PCR-based methods for detection of African lyssaviruses |
| title_full_unstemmed | Development of PCR-based methods for detection of African lyssaviruses |
| title_short | Development of PCR-based methods for detection of African lyssaviruses |
| title_sort | development of pcr based methods for detection of african lyssaviruses |
| topic | African lyssaviruses Polymerase chain reaction (PCR) Pcr-based methods UCTD |
| url | http://hdl.handle.net/2263/28545 http://upetd.up.ac.za/thesis/available/etd-10082010-143715/ |