Development of a serological assay for Chikungunya virus / Nishakanthi a/p Gopalan

Chikungunya virus (CHIKV) is a RNA virus that belongs to the Alphavirus genus of the family Togaviridae. It is transmitted by Aedes albopictus and Aedes aegypti, and causes clinical symptoms like fever with acute fever, skin rash and athralgia which mimics dengue fever. With current outbreaks of CHI...

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Main Author: Gopalan, Nishakanthi
Format: Thesis
Published: 2008
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Online Access:http://studentsrepo.um.edu.my/4373/1/Development_of_Diagnostic_Assay_of_Chikungunya_Virus_by_Nishakanthi_Gopalan.pdf
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author Gopalan, Nishakanthi
author_facet Gopalan, Nishakanthi
author_sort Gopalan, Nishakanthi
building UM Library
collection Institutional Repository
content_provider Universiti Malaya
content_source UM Student Repository
continent Asia
country Malaysia
description Chikungunya virus (CHIKV) is a RNA virus that belongs to the Alphavirus genus of the family Togaviridae. It is transmitted by Aedes albopictus and Aedes aegypti, and causes clinical symptoms like fever with acute fever, skin rash and athralgia which mimics dengue fever. With current outbreaks of CHIKV in Malaysia, it is essential to develop a serological assay as a tool for laboratory diagnosis and seroprevalence study. An IgG indirect ELISA was developed and used to validate nine samples collected from Bagan Panchor residents one year after an outbreak occurred in 2006. A variety of factors like varying cell culture types (C6/36 and Vero) to prepare the virus lysate antigen, different serum dilutions and the cut-off value determination methods were studied to optimize an IgG indirect ELISA assay. Neutralization assay was used as the gold standard. The IgG indirect ELISA using CHIKV-infected Vero cell lysate antigen performed better compared to CHIKV-infected C6/36 cell lysate antigen. The developed assay had a sensitivity of 100%, poor specificity of 25%, positive predictive value of 62.5%, negative predictive value of 100%, and concordance of 66.7%, compared to neutralization. The poor specificity and non-specific background readings are likely due to the crude total cell lysate used in the assay. Western blot identified the capsid protein as the immunogenic protein, which maybe used as a CHIKV recombinant antigen for further development of a more specific ELISA assay.
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spelling my.um.stud-43732014-09-27T01:35:02Z Development of a serological assay for Chikungunya virus / Nishakanthi a/p Gopalan Gopalan, Nishakanthi TP Chemical technology Chikungunya virus (CHIKV) is a RNA virus that belongs to the Alphavirus genus of the family Togaviridae. It is transmitted by Aedes albopictus and Aedes aegypti, and causes clinical symptoms like fever with acute fever, skin rash and athralgia which mimics dengue fever. With current outbreaks of CHIKV in Malaysia, it is essential to develop a serological assay as a tool for laboratory diagnosis and seroprevalence study. An IgG indirect ELISA was developed and used to validate nine samples collected from Bagan Panchor residents one year after an outbreak occurred in 2006. A variety of factors like varying cell culture types (C6/36 and Vero) to prepare the virus lysate antigen, different serum dilutions and the cut-off value determination methods were studied to optimize an IgG indirect ELISA assay. Neutralization assay was used as the gold standard. The IgG indirect ELISA using CHIKV-infected Vero cell lysate antigen performed better compared to CHIKV-infected C6/36 cell lysate antigen. The developed assay had a sensitivity of 100%, poor specificity of 25%, positive predictive value of 62.5%, negative predictive value of 100%, and concordance of 66.7%, compared to neutralization. The poor specificity and non-specific background readings are likely due to the crude total cell lysate used in the assay. Western blot identified the capsid protein as the immunogenic protein, which maybe used as a CHIKV recombinant antigen for further development of a more specific ELISA assay. 2008 Thesis NonPeerReviewed application/pdf http://studentsrepo.um.edu.my/4373/1/Development_of_Diagnostic_Assay_of_Chikungunya_Virus_by_Nishakanthi_Gopalan.pdf http://pendeta.um.edu.my/client/default/search/detailnonmodal/ent:$002f$002fSD_ILS$002f796$002fSD_ILS:796986/one?qu=Development+of+a+serological+assay+for+chikungunya+virus Gopalan, Nishakanthi (2008) Development of a serological assay for Chikungunya virus / Nishakanthi a/p Gopalan. Masters thesis, University of Malaya. http://studentsrepo.um.edu.my/4373/
spellingShingle TP Chemical technology
Gopalan, Nishakanthi
Development of a serological assay for Chikungunya virus / Nishakanthi a/p Gopalan
title Development of a serological assay for Chikungunya virus / Nishakanthi a/p Gopalan
title_full Development of a serological assay for Chikungunya virus / Nishakanthi a/p Gopalan
title_fullStr Development of a serological assay for Chikungunya virus / Nishakanthi a/p Gopalan
title_full_unstemmed Development of a serological assay for Chikungunya virus / Nishakanthi a/p Gopalan
title_short Development of a serological assay for Chikungunya virus / Nishakanthi a/p Gopalan
title_sort development of a serological assay for chikungunya virus / nishakanthi a/p gopalan
topic TP Chemical technology
url http://studentsrepo.um.edu.my/4373/1/Development_of_Diagnostic_Assay_of_Chikungunya_Virus_by_Nishakanthi_Gopalan.pdf
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url_provider http://studentsrepo.um.edu.my/