A diagnostic method based on MGB probes for rapid detection and simultaneous differentiation between virulent and vaccine strains of avian paramyxovirus type 1.
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.
Fratnik Steyer
Adela
A
Virology Unit, Institute for Microbiology and Parasitology, Veterinary Faculty, University of Ljubljana, Gerbiceva 60, SI-1115 Ljubljana, Slovenia. adela.fratnik@vf.uni-lj.si
Rojs
Olga Zorman
OZ
Krapez
Uros
U
Slavec
Brigita
B
Barlic-Maganja
Darja
D
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
D015345:Oligonucleotide Probes; D012367:RNA, Viral; D014765:Viral Vaccines