title: Real-time reverse transcription-polymerase chain reaction detection of Newcastle disease virus using light upon extension fluorogenic primers.
abstract: 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) EID(50)/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.
authors.affiliation: Central Veterinary Institute, Institute of Debrecen, Bornemissza u. 3-7., H-4031 Debrecen, Hungary.
authors.forename: Márta
authors.initials: M
authors.lastname: Antal
authors.forename: Tibor
authors.initials: T
authors.lastname: Farkas
authors.forename: Péter
authors.initials: P
authors.lastname: Germán
authors.forename: Sándor
authors.initials: S
authors.lastname: Belák
authors.forename: István
authors.initials: I
authors.lastname: Kiss
mesh_terms: D000818:Animals; D002645:Chickens / Q000821:virology; D017931:DNA Primers / Q000235:genetics*; D005243:Feces / Q000821:virology; D005456:Fluorescent Dyes; D005838:Genotype; D009522:Newcastle disease virus / Q000235:genetics* / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D014760:Viral Fusion Proteins / Q000235:genetics
chemical_list: D017931:DNA Primers; D005456:Fluorescent Dyes; D014760:Viral Fusion Proteins