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Primer, probe and reagent kit for RPA (recombinase polymerase amplification)-side flow chromatography detection of BRV (bovine rotavirus)

A technology of lateral flow chromatography and primer probes, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, microorganisms, etc. Increased sensitivity and ease of operation

Inactive Publication Date: 2018-09-18
SHANDONG NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the diagnosis methods of the disease mainly include pathogen isolation, PCR method and other technologies, because these methods require sophisticated instruments and cumbersome test procedures, it is difficult to meet the requirements of on-site detection in non-laboratory environments

Method used

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  • Primer, probe and reagent kit for RPA (recombinase polymerase amplification)-side flow chromatography detection of BRV (bovine rotavirus)
  • Primer, probe and reagent kit for RPA (recombinase polymerase amplification)-side flow chromatography detection of BRV (bovine rotavirus)
  • Primer, probe and reagent kit for RPA (recombinase polymerase amplification)-side flow chromatography detection of BRV (bovine rotavirus)

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Embodiment Construction

[0045] The present invention will be further described below in conjunction with the examples. It should be noted that the following descriptions are only for explaining the present invention and not limiting its content.

[0046] The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

[0047] TwistAmp nfo Kits were purchased from TwistDX Company, Genline Hybriddetect-1lateral flowstrips were purchased from Milenia GmbH (Germany), reverse transcription kit (Code No: D2639A) was purchased from Treasure Bioengineering (Dalian) Co., Ltd. DNA / RNA extraction of clinical samples Use Punch-it TM NA-Sample Kit (Nanohelix, Daejeon, South Korea); detection primers and probes were synthesized by Shanghai Sangon Biotechnology Co., Ltd.

[0048] The experimental method that does not indicate specific condition in the embodiment, generally according to routine condition, for example " molecular cloning: laboratory handb...

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Abstract

The invention discloses a primer, a probe and a reagent kit for RPA (recombinase polymerase amplification)-side flow chromatography detection of BRV (bovine rotavirus). The reagent kit contains primerprobe liquid for detecting the BRV through RPA technology, the forward primer sequence of the primer is as shown in SEQ ID No. 1, the reverse primer sequence is as shown in SEQ ID No. 2, and the probe sequence is as shown in SEQ ID No. 3. According to primer probe groups used in the invention, the RPA has a good amplification effect and strong strip specificity, does not have cross reaction withother viruses, has strong positive reaction in a detection area, and improves the sensitivity of detection. RPA-side flow chromatography technology is firstly adopted to establish a method for quick detection of the BRV, and by evaluation of specificity and sensitivity, the method can be used for clinical on-site detection, thereby providing a sensitive and reliable method for the on-site detection of the BRV.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer, a probe and a kit for detecting bovine rotavirus by applying recombinase polymerase amplification technology (Recombinase Polymerase Amplification, RPA) combined with lateral flow chromatography technology (Lateral Flow Assay) . Background technique [0002] Bovine rotavirus diarrhea is a diarrheal disease caused by rotavirus infection (Bovine rotavirus, BRV). It mainly infects calves aged 1 to 7 days. Diarrhea, dehydration and acid-base balance disturbance are the main clinical features. Since the intestinal cell damage caused by rotavirus is more conducive to the attachment and infection of pathogenic microorganisms such as toxin-producing Escherichia coli and Salmonella, this mixed infection and secondary infection will lead to more severe diarrhea and higher mortality. With the improvement of the intensive and large-scale scale of dairy cattle breeding in ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11C12R1/93
Inventor 何洪彬王洪梅贺文琦赵贵民侯佩莉
Owner SHANDONG NORMAL UNIV
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