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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
Online Access:http://psasir.upm.edu.my/id/eprint/8597/1/FSAS_1995_3_A.pdf
http://psasir.upm.edu.my/id/eprint/8597/
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spelling my.upm.eprints.85972012-05-07T09:17:04Z http://psasir.upm.edu.my/id/eprint/8597/ Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing 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. 1995 Thesis NonPeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/8597/1/FSAS_1995_3_A.pdf Ng, Ban Kim (1995) Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing. Masters thesis, Universiti Pertanian Malaysia. English
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
English
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
author Ng, Ban Kim
spellingShingle Ng, Ban Kim
Detection and Differentiation of Malaysian Newcastle Disease Virus Isolates by RNA-Polymerase Chain Reaction and Cycle Sequencing
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
publishDate 1995
url http://psasir.upm.edu.my/id/eprint/8597/1/FSAS_1995_3_A.pdf
http://psasir.upm.edu.my/id/eprint/8597/
_version_ 1643824063762661376
score 13.211869

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