Computational protocol: Simultaneous detection of eight immunosuppressive chicken viruses using a GeXP analyser-based multiplex PCR assay

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Protocol publication

[…] Gene-specific primers were designed based on sequence information obtained from GenBank using Primer premier 5.0 software (Premier, Palo Alto, USA) and NCBI Primer-Blast. ALV-A/B/J primers were designed to correspond to a specific region of the envelope gene gp85, MDV primers to a specific region of the gene encoding Marek’s EcoRI-Q protein (meq), REV primers to a specific region of the envelope gene gp90, IBDV primers to a specific region of the capsid protein gene VP2, CIAV primers to a specific region of the capsid protein gene VP1, and ARV primers to a specific region of the S1 gene. The primer sequences, the sizes of the resulting amplicons, and the target regions are listed in Table . The chimeric primers consisted of a universal sequence fused to the 5′-end of a gene-specific sequence. The forward universal primer was labelled at the 5′-end with the fluorescent dye Cy5. All chimeric primers and universal primers were synthesised and purified by polyacrylamide gel electrophoresis (Invitrogen, Shanghai, China). […]

Pipeline specifications

Software tools Primer Premier, Primer-BLAST
Application qPCR
Organisms Gallus gallus, Avian leukosis virus - RSA, Avian orthoreovirus
Diseases Marek Disease, HIV Infections, Coinfection