Differential amplification of Marek's disease CVI988 vaccine and of wild-type isolates from organs of commercial chickens using single or duplexed probes in real-time PCR

Avian Pathology : Journal of the W.V.P.A
I DavidsonYaad Dahan

Abstract

The differentiation of Marek's disease virus (MDV)-infected and vaccinated animal (DIVA) test, based on the MDV pp38 gene was described by Baigent et al. [(2016). Real-time PCR for differential quantification of CVI988 vaccine and virulent MDV strains. Journal of Virological Methods, 233, 23-36], using similar primers and alternate probes for virulent MDV-1 and the vaccine CVI988 virus. We explored the assay's applicability for commercial vaccines and commercial chickens, as the above-mentioned study employed tissue-cultured MDV strains and tissues from experimental trials. DNA of visceral organs and feathers of vaccinated or naturally infected chickens was used. Further, the applicability of the DIVA assay was evaluated using single or duplexed probes for the two viruses in the same amplification tube. Due to the high viral content in the commercial vaccines and in the clinical cases of MDV-1 infected commercial chickens, their examination by the MDV-1 DIVA real-time PCR was performed in one step. However, for the feather DNAs of commercially vaccinated birds, a step of pre-amplification was required. The MDV-1 DIVA real-time PCR performed as single probe in separate tubes using the Vir3 probe was very sensitive for virulent M...Continue Reading

References

May 1, 1978·Journal of the National Cancer Institute·K A Schat, B W Calnek
May 11, 2002·Journal of Veterinary Medicine. B, Infectious Diseases and Veterinary Public Health·I DavidsonY Weisman
Nov 23, 2005·Journal of Virological Methods·Mohamed Faizal Abdul-CareemShayan Sharif
Mar 22, 2007·The Journal of General Virology·Stephen J SpatzVenugopal Nair
Mar 15, 2016·Journal of Virological Methods·Susan J BaigentHervé Le Galludec

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Methods Mentioned

BETA
PCR

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DIVA

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