Evaluation of entamoeba histolytica acetyl co-a synthetase recombinant protein (rehacs) for diagnosis of acute amoebic liver abscess in moribund hamsters

Amoebic liver abscess (ALA) is a potentially fatal infection caused by Entamoeba histolytica. At Hospital Universiti Sains Malaysia, diagnosis of ALA includes detection of antibodies using a commercial kit based on crude soluble antigens (CSA) of E. histolytica. CSA consisted of a cocktail of E....

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Main Author:	Ching, Guee Cher
Format:	Thesis
Language:	English
Published:	2013
Subjects:	R Medicine
Online Access:	http://eprints.usm.my/54477/1/GUEE%20CHER%20CHING-24%20pages.pdf
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id	my-usm-ep.54477
record_format	uketd_dc
spelling	my-usm-ep.544772022-09-05T07:10:17Z Evaluation of entamoeba histolytica acetyl co-a synthetase recombinant protein (rehacs) for diagnosis of acute amoebic liver abscess in moribund hamsters 2013 Ching, Guee Cher R Medicine Amoebic liver abscess (ALA) is a potentially fatal infection caused by Entamoeba histolytica. At Hospital Universiti Sains Malaysia, diagnosis of ALA includes detection of antibodies using a commercial kit based on crude soluble antigens (CSA) of E. histolytica. CSA consisted of a cocktail of E. histolytica proteins that are not well-defmed in terms of the characteristics of the proteins and their masses. This study aimed to evaluate the diagnostic potential of a well-defined recombinant protein of E. histolytica acetyl-Co-A synthetase (rEhACS) using serum samples from hamsters with acute fatal ALA. The rEhACS was expressed in E. coli BL21 AI and purified using Ni-NTA resin column under optimized parameters. Purity of the protein was evaluated based on observation of the -77 kDa on SDS-PAGE gel stained with Coomassie brilliant blue. No unspecific band was observed in the gel. The specificity and sensitivity of the purified protein was evaluated via Western blot and enzyme-linked immunosorbent assay (ELISA) using 31 ALA-positive and 5 healthy hamster serum samples. Western blot analysis showed that the sensitivity and specificity ofrEhACS were 58.07% and 100% respectively; while the sensitivity and specificity of the recombinant EhACS used in an ELISA format were both 100%. In conclusion, the -77 kDa recombinant EhACS protein was shown to be important for diagnosis of acute fatal ALA in the animal model and its diagnostic potential in human ALA will be evaluated in future study. 2013 Thesis http://eprints.usm.my/54477/ http://eprints.usm.my/54477/1/GUEE%20CHER%20CHING-24%20pages.pdf application/pdf en public masters Universiti Sains Malaysia Pusat Pengajian Sains Perubatan
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	R Medicine
spellingShingle	R Medicine Ching, Guee Cher Evaluation of entamoeba histolytica acetyl co-a synthetase recombinant protein (rehacs) for diagnosis of acute amoebic liver abscess in moribund hamsters
description	Amoebic liver abscess (ALA) is a potentially fatal infection caused by Entamoeba histolytica. At Hospital Universiti Sains Malaysia, diagnosis of ALA includes detection of antibodies using a commercial kit based on crude soluble antigens (CSA) of E. histolytica. CSA consisted of a cocktail of E. histolytica proteins that are not well-defmed in terms of the characteristics of the proteins and their masses. This study aimed to evaluate the diagnostic potential of a well-defined recombinant protein of E. histolytica acetyl-Co-A synthetase (rEhACS) using serum samples from hamsters with acute fatal ALA. The rEhACS was expressed in E. coli BL21 AI and purified using Ni-NTA resin column under optimized parameters. Purity of the protein was evaluated based on observation of the -77 kDa on SDS-PAGE gel stained with Coomassie brilliant blue. No unspecific band was observed in the gel. The specificity and sensitivity of the purified protein was evaluated via Western blot and enzyme-linked immunosorbent assay (ELISA) using 31 ALA-positive and 5 healthy hamster serum samples. Western blot analysis showed that the sensitivity and specificity ofrEhACS were 58.07% and 100% respectively; while the sensitivity and specificity of the recombinant EhACS used in an ELISA format were both 100%. In conclusion, the -77 kDa recombinant EhACS protein was shown to be important for diagnosis of acute fatal ALA in the animal model and its diagnostic potential in human ALA will be evaluated in future study.
format	Thesis
qualification_level	Master's degree
author	Ching, Guee Cher
author_facet	Ching, Guee Cher
author_sort	Ching, Guee Cher
title	Evaluation of entamoeba histolytica acetyl co-a synthetase recombinant protein (rehacs) for diagnosis of acute amoebic liver abscess in moribund hamsters
title_short	Evaluation of entamoeba histolytica acetyl co-a synthetase recombinant protein (rehacs) for diagnosis of acute amoebic liver abscess in moribund hamsters
title_full	Evaluation of entamoeba histolytica acetyl co-a synthetase recombinant protein (rehacs) for diagnosis of acute amoebic liver abscess in moribund hamsters
title_fullStr	Evaluation of entamoeba histolytica acetyl co-a synthetase recombinant protein (rehacs) for diagnosis of acute amoebic liver abscess in moribund hamsters
title_full_unstemmed	Evaluation of entamoeba histolytica acetyl co-a synthetase recombinant protein (rehacs) for diagnosis of acute amoebic liver abscess in moribund hamsters
title_sort	evaluation of entamoeba histolytica acetyl co-a synthetase recombinant protein (rehacs) for diagnosis of acute amoebic liver abscess in moribund hamsters
granting_institution	Universiti Sains Malaysia
granting_department	Pusat Pengajian Sains Perubatan
publishDate	2013
url	http://eprints.usm.my/54477/1/GUEE%20CHER%20CHING-24%20pages.pdf
_version_	1747822284794494976

Evaluation of entamoeba histolytica acetyl co-a synthetase recombinant protein (rehacs) for diagnosis of acute amoebic liver abscess in moribund hamsters

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