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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| Format: | Article |
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Agricultural Research Council
2014
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| Online Access: | http://psasir.upm.edu.my/id/eprint/34955/ |
| _version_ | 1848847918329495552 |
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| author | Tan, Ching Giap Ideris, Aini Omar, Abdul Rahman Chen, Pei Yii Kleven, Stanley H. |
| author_facet | Tan, Ching Giap Ideris, Aini Omar, Abdul Rahman Chen, Pei Yii Kleven, Stanley H. |
| author_sort | Tan, Ching Giap |
| building | UPM Institutional Repository |
| collection | Online Access |
| 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. |
| first_indexed | 2025-11-15T09:26:14Z |
| format | Article |
| id | upm-34955 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-15T09:26:14Z |
| publishDate | 2014 |
| publisher | Agricultural Research Council |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-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 |
| 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 |
| title | 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_short | 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 |
| url | http://psasir.upm.edu.my/id/eprint/34955/ http://psasir.upm.edu.my/id/eprint/34955/ http://psasir.upm.edu.my/id/eprint/34955/ |