Computational protocol: Development of TaqMan® MGB fluorescent real-time PCR assay for the detection of anatid herpesvirus 1

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Protocol publication

[…] The FQ-PCR assay primers and probe (named Real-F, Real-R and Real-P respectively) design was carried out using the Primer Express™ software supplied by Applied Biosystems and their sequences were listed in Table . The forward and reverse primers amplified a 60 bp fragment of AHV-1 DNA polymerase gene as described (GenBank Accession No. AF064639). The fluorogenic probe was labelled at 5' with FAM (6-carboxyfluorescein) dye as reporter and labelled at 3' with TAMRA (tetra-methylcarboxyrhodamine) as quencher and 3'with MGB™ (Minor Groove Binder).The conventional PCR amplification was carried out using primers designed using the Primer Premier™ software according to the sequence as described (GenBank Accession No. AF064639). The forward primer and reverse primer (named Con-F and Con-R respectively) sequences were listed in Table and this primer pair yielded a 498 bp amplicon, in which the 60 bp FQ-PCR fragment was nested.All probes and primers were synthesized by Genecore Corporation (Shanghai, China) and purified by corresponding HPLC system. […]

Pipeline specifications

Software tools Primer Express, Primer Premier
Application qPCR
Organisms Anas platyrhynchos, Anatid alphaherpesvirus 1, Alcelaphine gammaherpesvirus 1
Diseases Infection