Newcastle disease

Farkas, T., Szekely, E., Belak, S., Kiss, I. 2009. Real-Time PCR-Base Pathotyping of Newcastle Disease Virus by Use of TaqMan Minor Groove Binder Probes.

• A protocol and primer set for reverse-transcription Real Time PCR detection of virulent and avirulent strains of Newcastle disease virus (NDV). RT-PCR targeted the fusion glycoprotein gene F0. This gene is somewhat variable allowing differentiation between virulent and avirulent strains but may result in false negatives in more variable strains. Primer sets were developed which targeted either lentogenic or mesogenic/velogenic strains. A TapMan MGB probe was also designed for each type to improve specificity. Of the other 24 pathogens tested with these primers none yielded false positives. RT-PCR by these methods had high specificity. Several sampling sites were tested and conjunctiva was found to be the best sample site.

Pham, H.M., Nakajima, C., Ohashi, K., Onuma, M. 2005. Loop-Mediated Isothermal Amplification for Rapid Detection of Newcastle Disease Virus.

• LAMP and PCR were used to detect NDV in experimentally infected chickens for several strains of NDV which included all degrees of virulence. The primers targeted the fusion gene F. Both PCR and LAMP were highly specific with no cross-reactivity for any of the three related viruses or three APMV serotypes tested. With LAMP, positive results were visualized equally well via electrophoresis gels and visual inspection of the reaction tube with added dye. Samples were taken from several tissues of infected chickens. Only samples from the lung and trachea yielded positive results; all other tissues resulted in false negatives.