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REAL-TIME PCR (qPCR ) WITH SYBR GREEN I TO DETECT DUCK VIRAL ENTERITIS (DVE)

Duck Virus Enteritis (DVE) is a viral disease which causes an acute contagious herpesvirus infection of waterfowl such as ducks, swans and geese and is a pathogen of economical concern. DVE diagnosis depends on histopathologic studies and serology, which are not effective in the early phase of the disease as it is laborious and time-consuming. As a result, a more effective and sensitive test was preferred. Although rapid methods such as conventional PCR and nested PCR are available, PCR-based methods are not carried to end point detection. Hence, a more rapid and sensitive end point assay is required. There are real time PCR methods which use Taqman and fluorogenic probe. Two qPCR systems were compared using GoTaq qPCR and PCR with SYBR green I based on melting temperatures discriminations. One specific melting peak was generated at temperatures 88.0±0.5°C represent DVE gene products. The real time PCR assay developed was sensitive where 10 DNA copies can be detected and DVE gene was detected specifically with no signal for other viruses such as AI, ND, CAV and EDS. Viral DNA could still be detected in infected cell cultures and field samples which were collected few years ago.

Keywords: Duck Viral Enteritis (DVE), real-time PCR, melting temperature, sensitive, specific.

Seng S.C., Syamsiah A.S., Khairul A.M., Maizan M., Ong G.H., Rozanah A.A.S. and Ramlan M.

Veterinary Research Institute, Department of Veterinary Services. 59 Jalan Sultan Azlan Shah, 31400 Ipoh, Perak.

Corresponding author: jesica_seng_168@yahoo.com

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Seng S.C., Syamsiah A.S., Khairul A.M., Maizan M., Ong G.H., Rozanah A.A.S. and Ramlan M. Real-Time PCR (qPCR) with SYBR Green I to Detect Duck Viral Enteritis (DVE). 24th VAM Congress 2012 (Proceedings p 314-317)-Poster  & MJVR-V2N1- pg17-25


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