Real-time fluorescence reverse transcription-polymerase chain reaction (RT-PCR) detection method and kit for highly pathogenic porcine reproductive and respiratory syndrome live vaccine

Abstract

The invention discloses nucleotide primer and probe sequences and a method for real-time fluorescence reverse transcription-polymerase chain reaction (RT-PCR) detection of a highly pathogenic porcine reproductive and respiratory syndrome vaccine, live (JXA1-R strain), and application of the method in detecting the highly pathogenic porcine reproductive and respiratory syndrome vaccine, live (JXA1-R strain). The method overcomes the defects of the traditional detection technology, has the characteristics of quickness, simplicity, convenience, strong specificity, high sensitivity and high reliability, and is suitable for quickly detecting the highly pathogenic porcine reproductive and respiratory syndrome vaccine, live (JXA1-R strain) and monitoring quality in vaccine production.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a real-time fluorescent RT-PCR detection method for highly pathogenic porcine reproductive and respiratory syndrome live vaccine (JXA1-R strain). Background technique [0002] Porcine reproductive and respiratory syndrome (Porcine reproductive and respiratory syndrome, PRRS) is caused by porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV). Characterized by highly contagious infectious diseases. The disease broke out in the United States for the first time in 1987, and then appeared in Canada, Europe and Asia one after another. At present, it has become popular all over the world, causing huge economic losses every year. PRRS first appeared in my country in 1995, and then spread rapidly. In 2006, there was an outbreak of highly pathogenic PRRSV (highly pathogenic PRRSV, HP-PRRSV) in China, which was characterized by high b...

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Problems solved by technology

[0003] Traditional PRRSV detection methods mainly include virus isolation and identification, immunoperoxidase monolayer assay (IPMA), indirect immunofluorescence assay (IFA), indirect enzyme-linked immunosorbent assay (indirect ELISA) and reverse transcription-polymerase Chain reaction test (RT-PCR), etc., they have played an important role in virus diagnosis and import and export inspection and quarantine, but each has shortcomings in sensitivity, specificity or timeliness, and it is difficult to isolate the vaccine strain (JXA1-R ) from wild strains to achieve the purpose of detecting vaccine strains (JXA1-R)

At present, although real-time fluorescent quantitative RT-PCR and other detection methods for highly pathogenic PRRSV variant strains have been established, they still cannot specifically detect the vaccine strain (JXA1-R)

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