Differentiation between pathogenic serotype 1 isolates of Marek's disease virus and the Rispens CVI988 vaccine in Australia using real-time PCR and high resolution melt curve analysis

Replication kinetics and shedding of very virulent Marek's disease virus and vaccinal Rispens/CVI988 virus during single and mixed infections varying in order and interval between infections

“…For determination of viral load various tissues, two TaqMan 1 qPCR assays were performed using a Rotor Gene 3000 realtime PCR instrument (Corbett Research, Australia). One test only detects Rispens/CVI988 MDV-1 while the complementary test only detects wild type pathogenic MDV-1 (Renz et al, 2013).…”

“…Recently Renz et al (2013) reported a qPCR method to differentiate between the Rispens virus and Australian isolates of pathogenic MDV-1. Based on the above discussion, we designed an experiment using this method to determine the effects of widely divergent vaccination to challenge intervals, including challenge prior to vaccination, on the level of vaccinal protection provided by the Rispens vaccine and the replication kinetics of Rispens and vvMDV in PBL, feather tips and dust samples.…”

A dCAPS assay detects and characterizes BaYMV according to its ability to overcome rym4-mediated resistance

“…Numerous SNP reading technologies have been described, such as High Resolution Melting analyses, SNaPshot and TaqMan real-time PCR assays (Balme-Sinibaldi et al, 2006;Renz et al, 2013;Rolland et al, 2008). However, the polymorphism of the sequences surrounding the targeted SNP does not necessarily allow the use of these techniques.…”

Detection of specific UL49 sequences of Marek's disease virus CVI988/Rispens strain using loop-mediated isothermal amplification

“…A huge step forward in detection and differentiation of MDV strain was achieved by the application of PCR and real-time PCR assays (Becker et al, 1992;Silva, 1992;Bumstead et al, 1997;Islam et al, 2004;Baigent et al, 2011). Using PCR-based techniques it was possible to differentiate all three MDV serotypes and also the attenuated CVI988/Rispens strain (Silva, 1992;Król et al, 2007;Baigent et al, 2011;Renz et al, 2013;Gimeno et al, 2014). Differentiation of the latter was exceptionally challenging as the differences between pathogenic MDV and Rispens strain are mainly located within genes encoding the 38 kDa phosphoprotein (pp38) (Cui et al, 1991;Silva, 1992;Reddy et al, 2002;Shamblin et al, 2004;Silva and Gimeno, 2007;Spatz et al, 2007a;Gimeno et al, 2014), immediately-early promoter (ICP4) (Spatz et al, 2012), and the repeated region of 132 bp sequence (Becker et al, 1992;Król et al, 2007;Silva et al, 2007).…”

“…Baigent et al (2005) showed that real-time PCR targeting meq gene is a convenient tool to monitor MDV load in feather tips. Also Islam et al (2004; and Renz et al (2013) applied real-time PCR to specifically detect Rispens in feather tips or dust. Recently, Gimeno et al (2014) showed that mismatch amplification mutation assay (MAMA) primers are capable to differentiate Rispens strain from the pathogenic form of the virus.…”