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Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum

The present study was based on the reverse transcription polymerase chain reaction (RT-PCR) of the 16S ribosomal nucleic acid (rRNA) of Mycoplasma for detection of viable Mycoplasma gallisepticum. To determine the stability of M. gallisepticum 16S rRNA in vitro, three inactivation methods were used...

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Main Authors: Tan, Ching Giap, Ideris, Aini, Omar, Abdul Rahman, Chen, Pei Yii, Kleven, Stanley H.
Format: Article
Published: Agricultural Research Council 2014
Online Access:http://psasir.upm.edu.my/id/eprint/34955/
http://www.ojvr.org/index.php/ojvr/article/view/708
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spelling my.upm.eprints.349552015-12-24T05:51:09Z http://psasir.upm.edu.my/id/eprint/34955/ Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum Tan, Ching Giap Ideris, Aini Omar, Abdul Rahman Chen, Pei Yii Kleven, Stanley H. The present study was based on the reverse transcription polymerase chain reaction (RT-PCR) of the 16S ribosomal nucleic acid (rRNA) of Mycoplasma for detection of viable Mycoplasma gallisepticum. To determine the stability of M. gallisepticum 16S rRNA in vitro, three inactivation methods were used and the suspensions were stored at different temperatures. The 16S rRNA of M. gallisepticum was detected up to approximately 20–25 h at 37 °C, 22–25 h at 16 °C, and 23–27 h at 4 °C. The test, therefore, could detect viable or recently dead M. gallisepticum (< 20 h). The RT-PCR method was applied during an in vivo study of drug efficacy under experimental conditions, where commercial broiler-breeder eggs were inoculated with M. gallisepticum into the yolk. Hatched chicks that had been inoculated in ovo were treated with Macrolide 1. The method was then applied in a flock of day 0 chicks with naturally acquired vertical transmission of M. gallisepticum, treated with Macrolide 2. Swabs of the respiratory tract were obtained for PCR and RT-PCR evaluations to determine the viability of M. gallisepticum. This study proved that the combination of both PCR and RT-PCR enables detection and differentiation of viable from non-viable M. gallisepticum. Agricultural Research Council 2014 Article PeerReviewed Tan, Ching Giap and Ideris, Aini and Omar, Abdul Rahman and Chen, Pei Yii and Kleven, Stanley H. (2014) Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum. Onderstepoort Journal of Veterinary Research, 81 (1). pp. 1-7. ISSN 0030-2465; ESSN: 2219-0635 http://www.ojvr.org/index.php/ojvr/article/view/708 10.4102/ojvr.v81i1.708
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/
description The present study was based on the reverse transcription polymerase chain reaction (RT-PCR) of the 16S ribosomal nucleic acid (rRNA) of Mycoplasma for detection of viable Mycoplasma gallisepticum. To determine the stability of M. gallisepticum 16S rRNA in vitro, three inactivation methods were used and the suspensions were stored at different temperatures. The 16S rRNA of M. gallisepticum was detected up to approximately 20–25 h at 37 °C, 22–25 h at 16 °C, and 23–27 h at 4 °C. The test, therefore, could detect viable or recently dead M. gallisepticum (< 20 h). The RT-PCR method was applied during an in vivo study of drug efficacy under experimental conditions, where commercial broiler-breeder eggs were inoculated with M. gallisepticum into the yolk. Hatched chicks that had been inoculated in ovo were treated with Macrolide 1. The method was then applied in a flock of day 0 chicks with naturally acquired vertical transmission of M. gallisepticum, treated with Macrolide 2. Swabs of the respiratory tract were obtained for PCR and RT-PCR evaluations to determine the viability of M. gallisepticum. This study proved that the combination of both PCR and RT-PCR enables detection and differentiation of viable from non-viable M. gallisepticum.
format Article
author Tan, Ching Giap
Ideris, Aini
Omar, Abdul Rahman
Chen, Pei Yii
Kleven, Stanley H.
spellingShingle Tan, Ching Giap
Ideris, Aini
Omar, Abdul Rahman
Chen, Pei Yii
Kleven, Stanley H.
Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum
author_facet Tan, Ching Giap
Ideris, Aini
Omar, Abdul Rahman
Chen, Pei Yii
Kleven, Stanley H.
author_sort Tan, Ching Giap
title Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum
title_short Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum
title_full Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum
title_fullStr Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum
title_full_unstemmed Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum
title_sort polymerase chain reaction-based discrimination of viable from non-viable mycoplasma gallisepticum
publisher Agricultural Research Council
publishDate 2014
url http://psasir.upm.edu.my/id/eprint/34955/
http://www.ojvr.org/index.php/ojvr/article/view/708
_version_ 1643831309656653824
score 13.211869

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