Optimization of viral RNA extraction methods for dengue virus detection in field collected Aedes albopictus / Nurul Hidayah Roslan

Dengue transmission has become a global threat and a health concern in tropical and sub-tropical countries. Aedes albopictus (Skuse), has high infestation rates in both urban and suburban areas of human settlement and is endemic in many Asian countries including Malaysia. The durability of the dengu...

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Main Author:	Roslan, Nurul Hidayah
Format:	Thesis
Language:	English
Published:	2015
Subjects:	Clinical pathology Laboratory technique
Online Access:	https://ir.uitm.edu.my/id/eprint/28103/1/TD_NURUL%20HIDAYAH%20ROSLAN%20HS%2015_5.pdf
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id	my-uitm-ir.28103
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spelling	my-uitm-ir.281032020-02-03T02:12:49Z Optimization of viral RNA extraction methods for dengue virus detection in field collected Aedes albopictus / Nurul Hidayah Roslan 2015 Roslan, Nurul Hidayah Clinical pathology. Laboratory technique Dengue transmission has become a global threat and a health concern in tropical and sub-tropical countries. Aedes albopictus (Skuse), has high infestation rates in both urban and suburban areas of human settlement and is endemic in many Asian countries including Malaysia. The durability of the dengue virus (DENV) has been correlated with the occurrence of transovarial transmission via its vector. The detection of transovarial DENV transmission is complex and requires high quality and concentration of viral RNA. There are many methods of extraction available in literature, some superior than others. The purpose of this study was to determine the most optimum extraction method for DENV detection and to detect transovarial transmission ofDENV using real time PCR (qRT-PCR) in field collected Ae. albopictus. A laboratory strain from USM was used to optimize the method of viral RNA extraction. Ae. Albopictus samples were collected from non-hotspot areas at Kolej Angsana, A5, UiTM Puncak Alam. The mosquitos were reared and hatched until adulthood in the laboratory. Both male and female Ae. Albopictus were selected, pooled and homogenized for RNA extraction. Modified PBS extraction with QIAamp viral RNA mini kit extraction protocol was then chosen for the sample extraction for this study. The outcome of this study has shown that the most suitable extraction method for DENV detection is by the use of modified PBS extraction method with QIAamp viral mini kit. In addition, no DENV was detected in all samples tested despite having optimized the best method for viral RNA extraction. This could be a result of the field work being collected during non-outbreak seasons, especially at non-hotspot areas. Further study is warranted to confirm these findings using a different locality. 2015 Thesis https://ir.uitm.edu.my/id/eprint/28103/ https://ir.uitm.edu.my/id/eprint/28103/1/TD_NURUL%20HIDAYAH%20ROSLAN%20HS%2015_5.pdf text en public degree Universiti Teknologi MARA Faculty of Health Sciences
institution	Universiti Teknologi MARA
collection	UiTM Institutional Repository
language	English
topic	Clinical pathology Laboratory technique
spellingShingle	Clinical pathology Laboratory technique Roslan, Nurul Hidayah Optimization of viral RNA extraction methods for dengue virus detection in field collected Aedes albopictus / Nurul Hidayah Roslan
description	Dengue transmission has become a global threat and a health concern in tropical and sub-tropical countries. Aedes albopictus (Skuse), has high infestation rates in both urban and suburban areas of human settlement and is endemic in many Asian countries including Malaysia. The durability of the dengue virus (DENV) has been correlated with the occurrence of transovarial transmission via its vector. The detection of transovarial DENV transmission is complex and requires high quality and concentration of viral RNA. There are many methods of extraction available in literature, some superior than others. The purpose of this study was to determine the most optimum extraction method for DENV detection and to detect transovarial transmission ofDENV using real time PCR (qRT-PCR) in field collected Ae. albopictus. A laboratory strain from USM was used to optimize the method of viral RNA extraction. Ae. Albopictus samples were collected from non-hotspot areas at Kolej Angsana, A5, UiTM Puncak Alam. The mosquitos were reared and hatched until adulthood in the laboratory. Both male and female Ae. Albopictus were selected, pooled and homogenized for RNA extraction. Modified PBS extraction with QIAamp viral RNA mini kit extraction protocol was then chosen for the sample extraction for this study. The outcome of this study has shown that the most suitable extraction method for DENV detection is by the use of modified PBS extraction method with QIAamp viral mini kit. In addition, no DENV was detected in all samples tested despite having optimized the best method for viral RNA extraction. This could be a result of the field work being collected during non-outbreak seasons, especially at non-hotspot areas. Further study is warranted to confirm these findings using a different locality.
format	Thesis
qualification_level	Bachelor degree
author	Roslan, Nurul Hidayah
author_facet	Roslan, Nurul Hidayah
author_sort	Roslan, Nurul Hidayah
title	Optimization of viral RNA extraction methods for dengue virus detection in field collected Aedes albopictus / Nurul Hidayah Roslan
title_short	Optimization of viral RNA extraction methods for dengue virus detection in field collected Aedes albopictus / Nurul Hidayah Roslan
title_full	Optimization of viral RNA extraction methods for dengue virus detection in field collected Aedes albopictus / Nurul Hidayah Roslan
title_fullStr	Optimization of viral RNA extraction methods for dengue virus detection in field collected Aedes albopictus / Nurul Hidayah Roslan
title_full_unstemmed	Optimization of viral RNA extraction methods for dengue virus detection in field collected Aedes albopictus / Nurul Hidayah Roslan
title_sort	optimization of viral rna extraction methods for dengue virus detection in field collected aedes albopictus / nurul hidayah roslan
granting_institution	Universiti Teknologi MARA
granting_department	Faculty of Health Sciences
publishDate	2015
url	https://ir.uitm.edu.my/id/eprint/28103/1/TD_NURUL%20HIDAYAH%20ROSLAN%20HS%2015_5.pdf
_version_	1783734053110808576

Optimization of viral RNA extraction methods for dengue virus detection in field collected Aedes albopictus / Nurul Hidayah Roslan

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