Rapid detection of Newcastle disease virus (NDV) with SYBR Green I real-time polymerase chain reaction
A two-step real-time PCR assay for the detection of Newcastle disease virus (NDV) was developed. Twelve NDV isolates (7 velogenic, 1 mesogenic and 4 lentogenic strains) were used. All the isolates showed positive results in amplification. Specificity of the amplification was confirmed by melting cur...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Conference or Workshop Item |
| Language: | en |
| Published: |
Universiti Putra Malaysia Press
2004
|
| Online Access: | http://psasir.upm.edu.my/id/eprint/65111/1/71-20.pdf http://psasir.upm.edu.my/id/eprint/65111/ |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832526682736558080 |
|---|---|
| author | Tan, Sheau Wei Ideris, Aini Omar, Abdul Rahman Mohd Yusoff, Khatijah Tan, Wen Siang |
| author_facet | Tan, Sheau Wei Ideris, Aini Omar, Abdul Rahman Mohd Yusoff, Khatijah Tan, Wen Siang |
| author_sort | Tan, Sheau Wei |
| building | UPM Library |
| collection | Institutional Repository |
| content_provider | Universiti Putra Malaysia |
| content_source | UPM Institutional Repository |
| continent | Asia |
| country | Malaysia |
| description | A two-step real-time PCR assay for the detection of Newcastle disease virus (NDV) was developed. Twelve NDV isolates (7 velogenic, 1 mesogenic and 4 lentogenic strains) were used. All the isolates showed positive results in amplification. Specificity of the amplification was confirmed by melting curve analysis. The melting temperatures (Tm) of all the isolates were between 86ºC to 87ºC. No primer dimer and non-specific products were detected during the amplification. A serial 10-fold dilution of cDNA was carried out to determine the sensitivity of the assay. In the optimal conditions, the detection limit of the real-time PCR was 10 pg whereas, the RT-nested ELISA PCR and conventional PCR can only detect up to 1 ng and 10 ng, respectively. Thus, SYBR Green I based real-time PCR assay offers a sensitive, rapid and convenient method in diagnosing NDV. |
| format | Conference or Workshop Item |
| id | my.upm.eprints-65111 |
| institution | Universiti Putra Malaysia |
| language | en |
| publishDate | 2004 |
| publisher | Universiti Putra Malaysia Press |
| record_format | eprints |
| spelling | my.upm.eprints-651112018-09-04T04:01:42Z http://psasir.upm.edu.my/id/eprint/65111/ Rapid detection of Newcastle disease virus (NDV) with SYBR Green I real-time polymerase chain reaction Tan, Sheau Wei Ideris, Aini Omar, Abdul Rahman Mohd Yusoff, Khatijah Tan, Wen Siang A two-step real-time PCR assay for the detection of Newcastle disease virus (NDV) was developed. Twelve NDV isolates (7 velogenic, 1 mesogenic and 4 lentogenic strains) were used. All the isolates showed positive results in amplification. Specificity of the amplification was confirmed by melting curve analysis. The melting temperatures (Tm) of all the isolates were between 86ºC to 87ºC. No primer dimer and non-specific products were detected during the amplification. A serial 10-fold dilution of cDNA was carried out to determine the sensitivity of the assay. In the optimal conditions, the detection limit of the real-time PCR was 10 pg whereas, the RT-nested ELISA PCR and conventional PCR can only detect up to 1 ng and 10 ng, respectively. Thus, SYBR Green I based real-time PCR assay offers a sensitive, rapid and convenient method in diagnosing NDV. Universiti Putra Malaysia Press 2004 Conference or Workshop Item PeerReviewed text en http://psasir.upm.edu.my/id/eprint/65111/1/71-20.pdf Tan, Sheau Wei and Ideris, Aini and Omar, Abdul Rahman and Mohd Yusoff, Khatijah and Tan, Wen Siang (2004) Rapid detection of Newcastle disease virus (NDV) with SYBR Green I real-time polymerase chain reaction. In: 11th International Conference of the Association of Institutions for Tropical Veterinary Medicine and 16th Veterinary Association Malaysia Congress, 23-27 Aug. 2004, Sunway Pyramid Convention Centre, Petaling Jaya, Malaysia. (pp. 240-241). |
| spellingShingle | Tan, Sheau Wei Ideris, Aini Omar, Abdul Rahman Mohd Yusoff, Khatijah Tan, Wen Siang Rapid detection of Newcastle disease virus (NDV) with SYBR Green I real-time polymerase chain reaction |
| title | Rapid detection of Newcastle disease virus (NDV) with SYBR Green I real-time polymerase chain reaction |
| title_full | Rapid detection of Newcastle disease virus (NDV) with SYBR Green I real-time polymerase chain reaction |
| title_fullStr | Rapid detection of Newcastle disease virus (NDV) with SYBR Green I real-time polymerase chain reaction |
| title_full_unstemmed | Rapid detection of Newcastle disease virus (NDV) with SYBR Green I real-time polymerase chain reaction |
| title_short | Rapid detection of Newcastle disease virus (NDV) with SYBR Green I real-time polymerase chain reaction |
| title_sort | rapid detection of newcastle disease virus (ndv) with sybr green i real-time polymerase chain reaction |
| url | http://psasir.upm.edu.my/id/eprint/65111/1/71-20.pdf http://psasir.upm.edu.my/id/eprint/65111/ |
| url_provider | http://psasir.upm.edu.my/ |