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RPA (Recombinase Ploymerase Amplification)-lateral flow assay detection primer of BPIV3 (Bovine parainfluenza virus type 3), probe and kit

A lateral flow chromatography, primer-probe technology, which is used in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of cumbersome test procedures, difficult to meet on-site detection, etc., and achieve good amplification effect. , the detection speed is fast, the effect of increasing the sensitivity

Inactive Publication Date: 2018-07-24
SHANDONG NORMAL UNIV
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  • Abstract
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Problems solved by technology

At present, the diagnostic methods of the disease mainly include pathogen isolation, PCR method, ELISA test and serum neutralization test, etc., 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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  • RPA (Recombinase Ploymerase Amplification)-lateral flow assay detection primer of BPIV3 (Bovine parainfluenza virus type 3), probe and kit
  • RPA (Recombinase Ploymerase Amplification)-lateral flow assay detection primer of BPIV3 (Bovine parainfluenza virus type 3), probe and kit
  • RPA (Recombinase Ploymerase Amplification)-lateral flow assay detection primer of BPIV3 (Bovine parainfluenza virus type 3), probe and kit

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Embodiment Construction

[0048] 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.

[0049] The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

[0050] 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 of clinical samples Extract using Punch-it TM NA-Sample Kit (Nanohelix, Daejeon, South Korea); detection primers and probes were synthesized by Shanghai Sangon Biotechnology Co., Ltd. The experimental method that does not indicate specific condition in the embodiment, generally according to routine condition, for example " molecular cloning: laboratory handbook " (Ne...

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Abstract

The invention discloses a primer and a probe for detecting BPIV3 (Bovine parainfluenza virus type 3) aerosol through a RPA (Recombinase Ploymerase Amplification)-lateral flow assay, a kit and a detection method. The kit contains primer probe liquid formed by a RPA primer and a probe for detecting the BPIV3, wherein the forward primer sequence is shown as SEQ ID No.1; the reverse primer sequence isshown as SEQ ID No.2; the probe sequence is shown as SEQ ID No.3. The used primer probe group has a good RPA effect; the strip specificity is high; no cross reaction with other viruses exists; strongpositive reaction exists in a detection region; the detection sensitivity is enhanced. The RPA-lateral flow assay technology is used for building a BPIV3 fast detection method for the first time; through specificity and sensitivity evaluation, the method can be used for clinical field detection; a flexible and reliable novel method is provided for BPIV3 field detection.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer, a probe and a kit thereof for detecting BPIV3 using a recombinase polymerase amplification technique (Recombinase Polymerase Amplification, RPA) combined with a lateral flow chromatography technique (Lateral FlowAssay). Background technique [0002] Bovine parainfluenza, also known as transport fever, is an acute contagious respiratory infectious disease caused by Bovine parainfluenzavirus type 3 (BPIV3) virus. And intensive cattle farms with high stocking density. BPIV3 is also one of the more important pathogens of bovine respiratory disease complex (BRDC), which mainly invades the respiratory tract, causing the immune function of the body and the function of macrophages to decline, resulting in immunosuppression of infected animals, and then secondary bacterial or mycoplasma infection. Typical BRDC occurs, leading to severe bronchitis, bronchiolitis, and pne...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
Inventor 王洪梅何洪彬高玉伟赵贵民侯佩莉贾春涛
Owner SHANDONG NORMAL UNIV
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