The invention belongs to the technical field of
poultry disease detection, and particularly relates to a poultry immunosuppressive
disease joint detection method based on multiple fluorescent quantitative PCR (
Polymerase Chain Reaction). The method sequentially comprises the following steps: determining a detection target, designing a specific primer and a probe, preparing a positive standard substance, pre-treating a sample, extracting
nucleic acid, purifying a
nucleic acid sample, establishing a multiple fluorescent quantitative PCR
reaction system, setting a multiple fluorescent quantitative PCR reaction program, amplifying, and analyzing and judging a result. On the basis, by designing a
multiplex fluorescent quantitative PCR detection
system and the specific primer probe, the kit has the advantages that
synchronous detection of multiple poultry immunosuppressive pathogens is realized, multiple independent detection on the same sample is not needed, the operation process is simplified, the detection cost is reduced, and the waste of the sample is reduced; the problems that a traditional single
pathogen detection method is low in efficiency and high in cost are solved.