Genome sequencing of Malaysian Newcastle disease viruses using next generation sequencing

Molecular detection and characterization of NDV focuses primarily on partial sequencing of the Fusion (F) gene. Complete genome sequencing by using Sanger sequencing protocols need large quantities of starting materials and high resource demanding methods such as cloning to a vectors prior to the se...

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Main Authors: Satharasinghe, Dilan Amila, Tan, Sheau Wei, Yeap, Swee Keong, Munir, Muhammad, Ideris, Aini, Omar, Abdul Rahman
Format: Conference or Workshop Item
Language:English
Published: Faculty of Veterinary Medicine, Universiti Putra Malaysia 2015
Online Access:http://psasir.upm.edu.my/id/eprint/65013/1/PA-14.pdf
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author Satharasinghe, Dilan Amila
Tan, Sheau Wei
Yeap, Swee Keong
Munir, Muhammad
Ideris, Aini
Omar, Abdul Rahman
author_facet Satharasinghe, Dilan Amila
Tan, Sheau Wei
Yeap, Swee Keong
Munir, Muhammad
Ideris, Aini
Omar, Abdul Rahman
author_sort Satharasinghe, Dilan Amila
collection UPM
description Molecular detection and characterization of NDV focuses primarily on partial sequencing of the Fusion (F) gene. Complete genome sequencing by using Sanger sequencing protocols need large quantities of starting materials and high resource demanding methods such as cloning to a vectors prior to the sequence acquisition. In this study, we described the application of a simple, robust and less resource demanding methodology for whole genome sequencing of NDV by using Next Generation Sequencing (NGS) technology. This protocol was able to generate accurate consensus-level full genome sequences for 5 virulent NDVs obtained from outbreaks during 2004 to 2013. The developed method works successfully for a limited quantity of starting material and has a potential to generate the consensus level sequences within a regular high-throughput diagnostic environment. In addition, the described protocol supports parallel sequencing of 94 samples in a single sequencing run which eventually reduce considerable amount of sequencing cost per sample. Phylogenetic analysis and Pair wise Sequencing Comparison (PASC) of both partial F gene and complete genome sequences confirmed the isolates clustered with genotype VIIa or lineage 5a.
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spelling upm.eprints-650132018-09-03T04:41:51Z http://psasir.upm.edu.my/id/eprint/65013/ Genome sequencing of Malaysian Newcastle disease viruses using next generation sequencing Satharasinghe, Dilan Amila Tan, Sheau Wei Yeap, Swee Keong Munir, Muhammad Ideris, Aini Omar, Abdul Rahman Molecular detection and characterization of NDV focuses primarily on partial sequencing of the Fusion (F) gene. Complete genome sequencing by using Sanger sequencing protocols need large quantities of starting materials and high resource demanding methods such as cloning to a vectors prior to the sequence acquisition. In this study, we described the application of a simple, robust and less resource demanding methodology for whole genome sequencing of NDV by using Next Generation Sequencing (NGS) technology. This protocol was able to generate accurate consensus-level full genome sequences for 5 virulent NDVs obtained from outbreaks during 2004 to 2013. The developed method works successfully for a limited quantity of starting material and has a potential to generate the consensus level sequences within a regular high-throughput diagnostic environment. In addition, the described protocol supports parallel sequencing of 94 samples in a single sequencing run which eventually reduce considerable amount of sequencing cost per sample. Phylogenetic analysis and Pair wise Sequencing Comparison (PASC) of both partial F gene and complete genome sequences confirmed the isolates clustered with genotype VIIa or lineage 5a. Faculty of Veterinary Medicine, Universiti Putra Malaysia 2015 Conference or Workshop Item PeerReviewed text en http://psasir.upm.edu.my/id/eprint/65013/1/PA-14.pdf Satharasinghe, Dilan Amila and Tan, Sheau Wei and Yeap, Swee Keong and Munir, Muhammad and Ideris, Aini and Omar, Abdul Rahman (2015) Genome sequencing of Malaysian Newcastle disease viruses using next generation sequencing. In: World Veterinary Poultry Association (Malaysia Branch) and World's Poultry Science Association (Malaysia Branch) Scientific Conference 2015, 21-22 Sept. 2015, Kuala Lumpur Convention Centre, Kuala Lumpur, Malaysia. (pp. 110-112).
spellingShingle Satharasinghe, Dilan Amila
Tan, Sheau Wei
Yeap, Swee Keong
Munir, Muhammad
Ideris, Aini
Omar, Abdul Rahman
Genome sequencing of Malaysian Newcastle disease viruses using next generation sequencing
title Genome sequencing of Malaysian Newcastle disease viruses using next generation sequencing
title_full Genome sequencing of Malaysian Newcastle disease viruses using next generation sequencing
title_fullStr Genome sequencing of Malaysian Newcastle disease viruses using next generation sequencing
title_full_unstemmed Genome sequencing of Malaysian Newcastle disease viruses using next generation sequencing
title_short Genome sequencing of Malaysian Newcastle disease viruses using next generation sequencing
title_sort genome sequencing of malaysian newcastle disease viruses using next generation sequencing
url http://psasir.upm.edu.my/id/eprint/65013/1/PA-14.pdf
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