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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| Format: | Thesis |
| Language: | English |
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Universiti Malaysia Sarawak, (UNIMAS)
2015
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| Online Access: | http://ir.unimas.my/id/eprint/10880/ http://ir.unimas.my/id/eprint/10880/1/Ratna%20Sri%20Dewi%20ft.pdf |
| _version_ | 1848836882124767232 |
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| author | Ratna Sri Dewi, Sedi |
| author_facet | Ratna Sri Dewi, Sedi |
| author_sort | Ratna Sri Dewi, Sedi |
| building | UNIMAS Institutional Repository |
| collection | Online Access |
| 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. |
| first_indexed | 2025-11-15T06:30:49Z |
| format | Thesis |
| id | unimas-10880 |
| institution | Universiti Malaysia Sarawak |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T06:30:49Z |
| publishDate | 2015 |
| publisher | Universiti Malaysia Sarawak, (UNIMAS) |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | unimas-108802023-07-03T06:32:58Z http://ir.unimas.my/id/eprint/10880/ Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay 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 NonPeerReviewed text en http://ir.unimas.my/id/eprint/10880/1/Ratna%20Sri%20Dewi%20ft.pdf Ratna Sri Dewi, Sedi (2015) Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay. Masters thesis, Universiti Malaysia Sarawak, (UNIMAS). |
| spellingShingle | R Medicine (General) Ratna Sri Dewi, Sedi Development of a rapid diagnostic test for enterovirus-71 using an isothermal amplification based assay |
| title | 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_short | 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 |
| topic | R Medicine (General) |
| url | http://ir.unimas.my/id/eprint/10880/ http://ir.unimas.my/id/eprint/10880/1/Ratna%20Sri%20Dewi%20ft.pdf |