Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing

This study was undertaken to develop diagnostic tests for Newcastle disease virus (NDV) based on RNApolymerase chain re action (RNA-PCR) and cycle sequencing techniques as a supplement to the presently available tests. Two RNA-PCR cycle sequencing systems each targeted at the haemagglutinin-ne...

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Main Author:	Ng, Ban Kim
Format:	Thesis
Language:	English English
Published:	1995
Subjects:	Newcastle disease virus - Malaysia Polymerase chain reaction - Case studies
Online Access:	http://psasir.upm.edu.my/id/eprint/8597/1/FSAS_1995_3_A.pdf
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id	my-upm-ir.8597
record_format	uketd_dc
spelling	my-upm-ir.85972012-05-07T09:17:04Z Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing 1995 Ng, Ban Kim This study was undertaken to develop diagnostic tests for Newcastle disease virus (NDV) based on RNApolymerase chain re action (RNA-PCR) and cycle sequencing techniques as a supplement to the presently available tests. Two RNA-PCR cycle sequencing systems each targeted at the haemagglutinin-neuraminidase (HN) and fusion (F) genes were developed. RNA-PCR amplification of a 398 base pairs (bp) HN gene fragment was performed on total RNAs extracted from infected allantoic fluid of 9 Malaysian NDV field isolates, vaccine strain V4-UPM and velogenic strain AF2240. Sequence analysis over 113 bases in the amplified fragment showed variations among these isolates/strains. However, identical sequences were obtained from some of the field isolates within a particular pathotype and these were thought to be of the same strain of NDV. RNA-PCR amplification of a 242 bp F gene fragment was performed on total RNAs which were isolated from several types of tissues (spleen, brain and lung) from six non-vaccinated chickens which were challenged with the velogenic strain AF2240. The spleen was found to have the highest number of samples positive by PCR. Spleens from uninfected chickens as well as those from vaccinated and challenged chickens which were slaughtered were all PCR negatives. The challenged virus was thought to have been neutralised by the antibodies produced by the chickens. The identity of the PCR products amplified from the spleens w'ere confirmed by cycle sequencing. Similarly, this test was also performed on RNAs from infected allantoic fluid of 11 different NDV strains/isolates. These samples were also PCR positives. Newcastle disease virus - Malaysia Polymerase chain reaction - Case studies 1995 Thesis http://psasir.upm.edu.my/id/eprint/8597/ http://psasir.upm.edu.my/id/eprint/8597/1/FSAS_1995_3_A.pdf application/pdf en public masters Universiti Pertanian Malaysia Newcastle disease virus - Malaysia Polymerase chain reaction - Case studies Faculty of Environmental Studies English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Newcastle disease virus - Malaysia Polymerase chain reaction - Case studies
spellingShingle	Newcastle disease virus - Malaysia Polymerase chain reaction - Case studies Ng, Ban Kim Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing
description	This study was undertaken to develop diagnostic tests for Newcastle disease virus (NDV) based on RNApolymerase chain re action (RNA-PCR) and cycle sequencing techniques as a supplement to the presently available tests. Two RNA-PCR cycle sequencing systems each targeted at the haemagglutinin-neuraminidase (HN) and fusion (F) genes were developed. RNA-PCR amplification of a 398 base pairs (bp) HN gene fragment was performed on total RNAs extracted from infected allantoic fluid of 9 Malaysian NDV field isolates, vaccine strain V4-UPM and velogenic strain AF2240. Sequence analysis over 113 bases in the amplified fragment showed variations among these isolates/strains. However, identical sequences were obtained from some of the field isolates within a particular pathotype and these were thought to be of the same strain of NDV. RNA-PCR amplification of a 242 bp F gene fragment was performed on total RNAs which were isolated from several types of tissues (spleen, brain and lung) from six non-vaccinated chickens which were challenged with the velogenic strain AF2240. The spleen was found to have the highest number of samples positive by PCR. Spleens from uninfected chickens as well as those from vaccinated and challenged chickens which were slaughtered were all PCR negatives. The challenged virus was thought to have been neutralised by the antibodies produced by the chickens. The identity of the PCR products amplified from the spleens w'ere confirmed by cycle sequencing. Similarly, this test was also performed on RNAs from infected allantoic fluid of 11 different NDV strains/isolates. These samples were also PCR positives.
format	Thesis
qualification_level	Master's degree
author	Ng, Ban Kim
author_facet	Ng, Ban Kim
author_sort	Ng, Ban Kim
title	Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing
title_short	Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing
title_full	Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing
title_fullStr	Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing
title_full_unstemmed	Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing
title_sort	detection and differentiation of malaysian newcastle disease virus isolates by rna-polymerase chain reaction and cycle sequencing
granting_institution	Universiti Pertanian Malaysia
granting_department	Faculty of Environmental Studies
publishDate	1995
url	http://psasir.upm.edu.my/id/eprint/8597/1/FSAS_1995_3_A.pdf
_version_	1747810835261030400

Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing

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