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

A technology for bovine viral diarrhea and bovine rotavirus, applied in recombinant DNA technology, biochemical equipment and methods, microbial assay/inspection, etc., can solve the problems of optimizing annealing temperature, difficulty, primer design and screening increase, etc. Achieve the effect of improving amplification efficiency, high sensitivity, and high sensitivity

Active Publication Date: 2019-11-08
NORTHEAST AGRICULTURAL UNIVERSITY
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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 ca

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  • 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
  • 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
  • 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

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Embodiment 1

[0041] Example 1 Establishment of multiple nano-PCR method for detection of bovine rotavirus, bovine parvovirus and bovine viral diarrhea virus based on DPO primers

[0042] 1. Design and synthesis of primers

[0043] 1.1 Design and synthesis of DPO primers for detection of bovine rotavirus

[0044] Use Oligo6.0 primer design software to design primers BRV-VP6-F and BRV-VP6-R according to the BRV NCDV VP6 gene sequence published in GenBank (accession number: JF693031.1) to amplify the full-length sequence of the VP6 gene (1172bp); On the basis of amplifying the full sequence of VP6, taking the VP6 gene as the target gene, selecting its conserved region to design DPO primers BRV-DPOF and BRV-DPOR and the corresponding conventional primers BRV-F and BRV-R (see table 1) The primers were synthesized by Shenggong Bioengineering (Shanghai) Co., Ltd.

[0045] Table 1 BRV primer sequence

[0046]

[0047] 1.2 Design and synthesis of DPO primers for detection of bovine parvovirus

[0048] Use O...

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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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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2537/143C12Q2563/137C12Q2563/155C12Q2545/113Y02A50/30
Inventor 乔薪瑗姜艳平崔文王丽周晗李一经唐丽杰徐义刚王孟孟
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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