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Journal of Veterinary Diagnostic Investigation Vol. 19 Issue 4, 400-404
Copyright © 2007 by the American Association of Veterinary Laboratory Diagnosticians
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Brief Communication

Real-time reverse transcription–polymerase chain reaction detection of Newcastle disease virus using light upon extension fluorogenic primers

Márta Antal, Tibor Farkas, Péter Germán, Sándor Belák and István Kiss1

Correspondence: 1Corresponding Author: István Kiss, Central Veterinary Institute, Institute of Debrecen, Bornemissza u. 3-7., H-4031 Debrecen, Hungary. istvan.kiss{at}sue.se

A real-time reverse transcriptase (RT)-PCR assay, applying light upon extension (LUX) fluorogenic primers, was developed for rapid and efficient detection of Newcastle disease virus (NDV). The method, which targets the fusion (F) protein gene of the viral genome, gave positive signal with all NDV isolates tested (32/32), while negative results were obtained with heterologous pathogens (35/35), including 13 avian influenza virus isolates. The detection limit of the assay was approximately 10+1.2 egg infectious dose (EID)50/0.2 ml and 10+2.2 EID50/0.2 ml for virus suspensions and spiked chicken fecal samples, respectively. As expressed in plasmid copy number, the procedure has a sensitivity of approximately 20 copies of the plasmid harboring the target gene. Due to its high specificity, sensitivity, and relative simplicity, the LUX RT-PCR assay provides a novel, rapid, and practical tool for the detection of NDV.

Key Words: Light upon extension (LUX) • Newcastle disease virus • real-time RT-PCR




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 J. Clin. Microbiol.Home page
T. Farkas, E. Szekely, S. Belak, and I. Kiss
Real-Time PCR-Based Pathotyping of Newcastle Disease Virus by Use of TaqMan Minor Groove Binder Probes
J. Clin. Microbiol., July 1, 2009; 47(7): 2114 - 2123.
[Abstract] [Full Text] [PDF]


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