20951166
not annotated - annotated - LINNAEUS only
Rapid detection of Newcastle disease virus replication in embryonated chicken eggs using quantitative real time polymerase chain reaction.
Newcastle disease virus (NDV), an avian paramyxovirus, is an economically important disease of poultry globally. Rapid methods to detect and differentiate the virus are important to curtail the spread of this virus. Nucleic acid based detection methods are routinely employed for diagnosis that suffer from the disadvantage of failure to discriminate viable virus and non-infectious genome. However, virus isolation remains the gold standard for diagnosis of field outbreaks. The sensitivity of virus isolation was combined with nucleic acid based detection methods so that the time taken for confirmatory diagnosis could be considerably reduced while increasing sensitivity. Quantitative real time reverse transcription polymerase chain reaction (qRT-PCR) and conventional RT-PCR techniques were compared for the detection of NDV genome replication in 9-11-day-old embryonated chicken eggs (ECE) using the nucleoprotein (NP) gene of the virus as a target. The results suggest that at least two to fourfold increase in cycle threshold (C(t)) values over the baseline C(t) value of samples lacking infectious virus, would indicate live NDV replication. The limit of detection of NDV replication using qRT-PCR was 1x10(4.0) mean embryo infective doses (EID(50)). The earliest time point when live virus replication was detectable by qRT-PCR or RT-PCR was 30h post-inoculation in ECE.
Ann file
T1 Species 19 42 Newcastle disease virus
N1 Reference T1 Taxonomy:11176
T2 Species 70 77 chicken
N2 Reference T2 Taxonomy:9031
T3 Species 141 164 Newcastle disease virus
N3 Reference T3 Taxonomy:11176
T4 Species 166 169 NDV
N4 Reference T4 Taxonomy:11176
T5 Out-of-scope 236 243 poultry
N5 Reference T5 Taxonomy:9031
T6 Species 968 971 NDV
N6 Reference T6 Taxonomy:11176
T7 Species 1019 1026 chicken
N7 Reference T7 Taxonomy:9031
T8 Species 1276 1279 NDV
N8 Reference T8 Taxonomy:11176
T9 Species 1319 1322 NDV
N9 Reference T9 Taxonomy:11176
#1 AnnotatorNotes T5 TODO check