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Double rpa detection kit for bovine parainfluenza virus and bovine respiratory syncytial virus

A technology of bovine parainfluenza virus and detection kit, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, biochemical equipment and methods, etc., can solve the problems that cannot be designed and realized, and have strict requirements, and achieve fast detection speed and low cost. Low, convenient and rapid screening effect

Active Publication Date: 2022-07-05
NORTHWEST A & F UNIV
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AI Technical Summary

Problems solved by technology

Compared with PCR, RPA has more stringent requirements on primer design, which cannot be realized completely according to software design. Screening effective specific amplification primers and reaction conditions is a difficult problem in realizing multiple RPA detection.

Method used

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  • Double rpa detection kit for bovine parainfluenza virus and bovine respiratory syncytial virus
  • Double rpa detection kit for bovine parainfluenza virus and bovine respiratory syncytial virus
  • Double rpa detection kit for bovine parainfluenza virus and bovine respiratory syncytial virus

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Embodiment Construction

[0035] The present invention will be described in further detail below with reference to the accompanying drawings and embodiments. The embodiments are only used to explain the present invention, but not to limit the protection scope of the present invention.

[0036] (1) Primer design and screening

[0037] (1) Collect the whole genome sequences of bovine parainfluenza virus type 3 (BPIV-3) and bovine respiratory syncytial virus (BRSV) in GeneBank, and use MegAlign to align the sequences (BPIV-3 is 12 The whole genome sequence, BRSV is 5 whole genomes), select the intra-species conserved and inter-species specific region as the detection target gene of each virus, determine the NP gene of BPIV-3 and the N gene of BRSV as target genes, and further compare The target gene sequence is determined, and the candidate region of the gene sequence to be amplified is determined, and then primers are designed according to the determined gene sequence. Among them, the RPA primer design...

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Abstract

The invention discloses a double RPA detection kit for bovine parainfluenza virus and bovine respiratory syncytial virus, comprising bovine parainfluenza virus composed of nucleotide sequences shown in SEQ.ID.NO.2 and SEQ.ID.NO.5 A primer set for RPA detection, and a primer set for bovine respiratory syncytial virus RPA detection consisting of the nucleotide sequences shown in SEQ.ID.NO.8 and SEQ.ID.NO.10. The kit is based on the recombinase polymerase amplification method, uses the cDNA formed by reverse transcription of RNA as a template, performs amplification reaction under optimized reaction temperature and time, and uses nucleic acid gel electrophoresis to obtain detection results. The invention can be used to detect bovine parainfluenza virus and bovine respiratory syncytial virus simultaneously, has simple operation, high sensitivity and specificity, and provides a rapid detection reagent for on-site screening of bovine pathogens.

Description

technical field [0001] The invention relates to the field of viral nucleic acid detection, in particular to a dual RPA detection primer set, kit and detection method for bovine parainfluenza virus and bovine respiratory syncytial virus. Background technique [0002] Bovine parainfluenza is a disease characterized by high fever, dyspnea and cough and mainly respiratory symptoms caused by bovine parainfluenza-3 virus (BPIV-3) infection. The virus appears mixed infection. BPIV-3 infection can cause respiratory function impairment and immunosuppression in cattle, and lead to death of cattle, which is extremely harmful to house-feeding and fattening cattle. Bovine respiratory syncytial virus (BRSV) is an enveloped, single-stranded, negative-strand RNA virus that can cause miscarriage and permanent lung damage after infection, promote opportunistic secondary pathogen invasion, and be harmful to animal production. long-term loss of performance. [0003] Most of the farms are loc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/701C12Q1/6844C12Q2600/16C12Q2531/119C12Q2537/143C12Q2521/507C12Q2537/1376C12Q2565/125Y02A50/30
Inventor 赵晓民童德文王凯丽刘丹
Owner NORTHWEST A & F UNIV
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