title: A diagnostic method based on MGB probes for rapid detection and simultaneous differentiation between virulent and vaccine strains of avian paramyxovirus type 1.
abstract: Newcastle disease virus (NDV), also designated avian paramyxovirus type 1 (APMV-1), is a serious pathogen of poultry, causing highly contagious Newcastle disease (ND), with high morbidity or mortality, depending on the strain. Accordingly, rapid and reliable detection of APMV-1 and differentiation between vaccine and virulent strains is of crucial importance for ND diagnosis and plays an important role in effective control of the disease. In this study, two real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assays using minor groove-binding (MGB) probes were developed for broad range detection and simultaneous pathotyping of APMV-1. The two assays were evaluated for their ability to detect in allantoic fluids viral RNA of all known APMV-1 lineages. Additionally, the applicability of the developed assays was assessed by the detection and pathotype prediction of APMV-1 in swabs and organs. The assays demonstrated high analytical specificity, sensitivity and good reproducibility, with coefficients of variation ranging from 0.2% to 3.9% and from 0.6% to 7.2% for intra-assay and inter-assay variability, respectively. The results indicated the suitability of both assays as a complementary method for rapid screening and typing of APMV-1.
authors.affiliation: Virology Unit, Institute for Microbiology and Parasitology, Veterinary Faculty, University of Ljubljana, Gerbiceva 60, SI-1115 Ljubljana, Slovenia. adela.fratnik@vf.uni-lj.si
authors.forename: Adela
authors.initials: A
authors.lastname: Fratnik Steyer
authors.forename: Olga Zorman
authors.initials: OZ
authors.lastname: Rojs
authors.forename: Uros
authors.initials: U
authors.lastname: Krapez
authors.forename: Brigita
authors.initials: B
authors.lastname: Slavec
authors.forename: Darja
authors.initials: D
authors.lastname: Barlic-Maganja
mesh_terms: D000825:Animal Structures / Q000821:virology; D000818:Animals; D018109:Avulavirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D001483:Base Sequence; D001715:Bird Diseases / Q000175:diagnosis* / Q000821:virology; D002645:Chickens; D010856:Columbidae; D008969:Molecular Sequence Data; D009521:Newcastle Disease / Q000175:diagnosis* / Q000821:virology; D015345:Oligonucleotide Probes / Q000235:genetics; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014765:Viral Vaccines
chemical_list: D015345:Oligonucleotide Probes; D012367:RNA, Viral; D014765:Viral Vaccines