Multi-nano-PCR primer group for detecting bovine rotavirus (BRV), bovine parvovirus (BPV) and bovine viral diarrhea virus (BVDV) and application of multi-nano-PCR primer group

Abstract

The invention discloses a multi-nano-PCR primer group for detecting bovine rotavirus (BRV), bovine parvovirus (BPV) and a bovine viral diarrhea virus (BVDV) and application of the multi-nano-PCR primer group. The primer group contains dual-priming oligonucleotide (DPO) primer pairs used for detecting the BRV, the BPV and the BVDV correspondingly. A multi-DPO-nano-PCR detection method capable of simultaneously detecting the BRV, the BPV and the BVDV is established by combining DPO primers with a nano PCR technology. Compared with a conventional PCR method, the multi-DPO-nano-PCR detection method is time-saving and labor-saving, can quickly, accurately and specifically detect pathogens, and achieves high sensitivity on the basis of ensuring specificity. By providing the multi-nano PCR primergroup, the new method is provided for diagnosis of early infection and inapparent infection of the BRV, the BPV and the BVDV, a reliable technical means is provided for detection of clinical mixed infection of the BRV, the BPV and the BVD, and technical support is provided for epidemic disease testing, screening purification and comprehensive prevention and control.

Description

Technical field [0001] The invention relates to a multiple PCR primer set for detecting bovine rotavirus, bovine parvovirus and bovine viral diarrhea virus and its application, in particular to a multiple PCR primer set for detecting bovine rotavirus, bovine parvovirus and bovine viral diarrhea virus The multiple DPO-nanoPCR primer set of DPO also relates to a nano PCR method for detecting bovine rotavirus, bovine parvovirus and bovine viral diarrhea virus based on the DPO primer set. The present invention belongs to the field of biotechnology. Background technique [0002] With the development of animal husbandry, there are more and more large-scale and intensive breeding, and the contact between cattle herds has increased. This often brings opportunities for the spread of contagious infectious diseases, and the speed of disease transmission accelerates, making the diseased cattle often There is a mixed infection of two or more pathogens. In 2007, the National Animal Health Mon...

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Problems solved by technology

[0004] At present, the diagnosis of BRV, BPV, and BVDV mainly includes electron microscopy, virus isolation, serological diagnosis and molecular biology methods. Virus isolation, electron microscopy and serological methods generally cannot detect multiple pathogens at the same time, and are not suitable for the diagnosis of mixed infections.

The multiplex PCR method is often used in the detection of mixed infections, which can amplify multiple target genes at the same time, which is convenient and fast. However, multiplex PCR technology needs to consider many factors when designing primers. Since the annealing temperature of multiple pairs of primers is similar, it should be avoided. Generate primer dimers and hairpin structures, and at the same time need to optimize the annealing temperature, which adds difficulties to primer design and screening

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