UPM Institutional Repository

Rapid detection of Newcastle disease virus (NDV) with SYBR Green I real-time polymerase chain reaction


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).


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.

Download File

[img] Text
Restricted to Repository staff only

Download (87kB)

Additional Metadata

Item Type: Conference or Workshop Item (Paper)
Divisions: Faculty of Science and Environmental Studies
Faculty of Veterinary Medicine
Publisher: Universiti Putra Malaysia Press
Keywords: Newcastle disease virus; SYBR Green I; Real time PCR; RT-nested PCR ELISA
Depositing User: Nabilah Mustapa
Date Deposited: 04 Sep 2018 04:01
Last Modified: 04 Sep 2018 04:01
URI: http://psasir.upm.edu.my/id/eprint/65111
Statistic Details: View Download Statistic

Actions (login required)

View Item View Item