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Dual RPA detection kit for bovine parainfluenza virus (BPIV) and bovine respiratory syncytial virus (BRSV)

A bovine parainfluenza virus and detection kit technology, applied in the direction of recombinant DNA technology, DNA / RNA fragments, biochemical equipment and methods, etc., can solve the problems of strict requirements and inability to design and realize, and achieve low cost and high detection speed Fast, sensitive and stable results

Active Publication Date: 2021-04-02
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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  • Dual RPA detection kit for bovine parainfluenza virus (BPIV) and bovine respiratory syncytial virus (BRSV)
  • Dual RPA detection kit for bovine parainfluenza virus (BPIV) and bovine respiratory syncytial virus (BRSV)
  • Dual RPA detection kit for bovine parainfluenza virus (BPIV) and bovine respiratory syncytial virus (BRSV)

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Embodiment Construction

[0035] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments. The examples are only used to explain the present invention, 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 compare and analyze the sequences (BPIV-3 is 12 The whole genome sequence, BRSV is 5 whole genomes), select the region that is conserved within the species and specific among the species as the detection target gene of each virus, determine the NP gene of BPIV-3 and the N gene of BRSV as the target gene, and further compare Target gene sequence, determine the candidate region of the gene sequence to be amplified, and then design primers based on the determined gene sequence. Among them, the RPA primer design of BPI...

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Abstract

The invention discloses a dual RPA detection kit for bovine parainfluenza virus (BPIV) and bovine respiratory syncytial virus (BRSV). The dual RPA detection kit comprises aBPIV RPA detection primer group consisting of nucleotide sequences shown in SEQ. ID. NO.2 and SEQ. ID. NO.5, and a BRSV RPA detection primer group consisting of nucleotide sequences shown in SEQ. ID. NO.8 and SEQ. ID. NO.10. Thekit is based on a recombinase polymerase amplification method, cDNA formed by RNA reverse transcription is used as a template, an amplification reaction is carried out at an optimized reaction temperature and time, and a detection result is obtained by nucleic acid gel electrophoresis. The kit can be used for simultaneously detecting the bBPIV and the BRSV, is simple to operate, has higher 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 caused by bovine parainfluenza virus type 3 (Bovine parainfluenza-3virus, BPIV-3) infection, which is characterized by high fever, dyspnea and cough and mainly respiratory symptoms, often associated with other respiratory symptoms. Virus mixed infection. BPIV-3 infection can cause respiratory function damage and immunosuppression in cattle, and lead to death of cattle. It is extremely harmful to fattening cattle in house feeding. It is considered to be the main pathogen of pneumonia that causes major economic losses in intensive farming around the world. Bovine respiratory syncytial virus (BRSV) is an enveloped, single-stranded, negative-sense RNA virus that can cause abortion and permane...

Claims

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Application Information

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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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