Detection of Newcastle disease virus using a SYBR Green I real time polymerase chain reaction

• Main Authors: Tan, Sheau Wei, Omar, Abdul Rahman, Ideris, Aini, Yusoff, Khatijah, Tan, Wen Siang
• Format: Article
• Language: English
• Published: AEPress 2004
• Online Access: http://psasir.upm.edu.my/id/eprint/692/1/PFBSB8.pdf; http://psasir.upm.edu.my/id/eprint/692/; http://www.elis.sk/index.php?page=shop.product_details&flypage=flypage.tpl&product_id=45&category_id=3&option=com_virtuemart&vmcchk=1&Itemid=1

A two-step SYBR Green I real time polymerase chain reaction (PCR, real time PCR) for the detection of Newcastle disease virus (NDV) was developed. A melting curve analysis was performed to distinguish specific from non-specific products and primer dimmers. Regardless of different virus pathotypes the melting temperature (Tm) ranged from 86°C to 87°C. The sensitivity of the real time PCR was compared with the reverse transcription (RT) – nested PCR enzyme – linked immunosorbent assay (ELISA, RT-nested PCR ELISA). Whereas the detection limit of the real time PCR was 10 pg DNA, the RT-nested PCR ELISA and conventional PCR could only detect up to 1 ng and DNA, respectively. Thus the real time PCR offers a sensitive, rapid and convenient method for screening large number of NDV specimens.