Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay

An in-house nucleic acid sequence based amplification (NASBA) assay protocol for the detection of EV-A71 from both virus isolates and clinical specimens was developed in this study. A primer set, targeting the VP1 gene region, was designed and the sensitivity and specificity was tested using conv...

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Main Author:	Ratna Sri Dewi, Sedi
Format:	Thesis
Language:	English
Published:	2015
Subjects:	R Medicine (General)
Online Access:	http://ir.unimas.my/id/eprint/10880/1/Ratna%20Sri%20Dewi%20ft.pdf
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spelling	my-unimas-ir.108802023-07-03T06:32:58Z Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay 2015 Ratna Sri Dewi, Sedi R Medicine (General) An in-house nucleic acid sequence based amplification (NASBA) assay protocol for the detection of EV-A71 from both virus isolates and clinical specimens was developed in this study. A primer set, targeting the VP1 gene region, was designed and the sensitivity and specificity was tested using conventional RT-PCR and subsequently used in a NASBA assay. The detection of EV-A71 NASBA amplicons was successfully achieved using ethidium bromide-stained RNA agarose gel electrophoresis. The in-house EV-A71 NASBA protocol detected a panel of 54 of 60 EV-A71 virus isolates from different genogroups and subgenogroups and no cross-reactivity was observed against other EV serotypes. The NASBA assay was additionally evaluated on a panel of, 41 clinical specimens of various sample types from HFMD cases in Sarawak. Twenty-one (95.5%) of 22 clinical specimens that were confirmed to be EV-A71 positive by RT-PCR and virus isolation were successfully detected by the NASBA assay. The other 19 clinical specimens that were concordantly EV-A71 negative by both RT-PCR and virus isolation, were not detected by the EV-A71 NASBA assay. Verification of the EV-A71 NASBA amplicons from virus isolates and clinical specimens were successfully achieved by sequencing of the amplified region. In conclusion, the NASBA assay developed in this study provides a useful alternative for the screening and detection of EV-A71. Universiti Malaysia Sarawak, (UNIMAS) 2015 Thesis http://ir.unimas.my/id/eprint/10880/ http://ir.unimas.my/id/eprint/10880/1/Ratna%20Sri%20Dewi%20ft.pdf text en validuser masters Universiti Malaysia Sarawak, (UNIMAS) Institute of Health and Communiti Medicine
institution	Universiti Malaysia Sarawak
collection	UNIMAS Institutional Repository
language	English
topic	R Medicine (General)
spellingShingle	R Medicine (General) Ratna Sri Dewi, Sedi Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay
description	An in-house nucleic acid sequence based amplification (NASBA) assay protocol for the detection of EV-A71 from both virus isolates and clinical specimens was developed in this study. A primer set, targeting the VP1 gene region, was designed and the sensitivity and specificity was tested using conventional RT-PCR and subsequently used in a NASBA assay. The detection of EV-A71 NASBA amplicons was successfully achieved using ethidium bromide-stained RNA agarose gel electrophoresis. The in-house EV-A71 NASBA protocol detected a panel of 54 of 60 EV-A71 virus isolates from different genogroups and subgenogroups and no cross-reactivity was observed against other EV serotypes. The NASBA assay was additionally evaluated on a panel of, 41 clinical specimens of various sample types from HFMD cases in Sarawak. Twenty-one (95.5%) of 22 clinical specimens that were confirmed to be EV-A71 positive by RT-PCR and virus isolation were successfully detected by the NASBA assay. The other 19 clinical specimens that were concordantly EV-A71 negative by both RT-PCR and virus isolation, were not detected by the EV-A71 NASBA assay. Verification of the EV-A71 NASBA amplicons from virus isolates and clinical specimens were successfully achieved by sequencing of the amplified region. In conclusion, the NASBA assay developed in this study provides a useful alternative for the screening and detection of EV-A71.
format	Thesis
qualification_level	Master's degree
author	Ratna Sri Dewi, Sedi
author_facet	Ratna Sri Dewi, Sedi
author_sort	Ratna Sri Dewi, Sedi
title	Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay
title_short	Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay
title_full	Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay
title_fullStr	Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay
title_full_unstemmed	Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay
title_sort	development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay
granting_institution	Universiti Malaysia Sarawak, (UNIMAS)
granting_department	Institute of Health and Communiti Medicine
publishDate	2015
url	http://ir.unimas.my/id/eprint/10880/1/Ratna%20Sri%20Dewi%20ft.pdf
_version_	1783728080636870656

Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay

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