RPA-LFA (recombinase polymerase amplification-lateral flow assay) detection kit for detecting brucella and use method of kit

A detection kit, Brucella technology, applied in the fields of medicine and biology, can solve the problems of difficult to meet the detection needs, expensive, long time, etc., and achieve the effect of lowering the technical threshold, easy operation, and broad market prospects

Inactive Publication Date: 2019-02-15
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above methods all require expensive laboratory conditions, equipment and professional skills, and take a long time, which is difficult to meet the detection needs under field or field conditions.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A kind of RPA-LFD detection kit that detects Brucella, comprises: (1) sample treatment liquid: the PBS solution of 0.1M pH7.4; (2) detection reaction liquid: the Tris-HCl of 50mM pH7.9; 100mM Potassium acetate; 2mM dithiothreitol; 3mM adenosine triphosphate; 20mM creatine phosphate; 5μg creatine kinase; 5% polyethylene glycol, 200μM deoxyribonucleoside triphosphate, primer (Primers BBRRPAF and BBRRPAB each 0.4μM, probe BBRRPAP 0.05μM), 14mM magnesium acetate; (3) dry powder tube: 3.75μg lyophilized recombinant enzyme uvsX, 750ng lyophilized recombinant enzyme uvsY, 30μg single-chain binding protein , 900ng strand displacement polymerase; (4) lateral flow test strip; (5) positive control reaction solution: 100 copies of the BvrR gene; TE buffer solution of 10mMpH8.0; (6) negative control reaction solution: 10mM pH8. 0 TE buffer solution; (7) loading treatment solution: PBST buffer solution of 0.01MpH7.3;

[0032] Among them, the forward primer is

[0033] BBRRPAF: CTCA...

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PUM

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Abstract

The invention discloses an RPA-LFA (recombinase polymerase amplification-lateral flow assay) detection kit for detecting brucella. The kit comprises (1) a sample tretament solution, (2) a detection reaction solution, (3) a dry powder tube, (4) a lateral flow test strip, (5) a positive control reaction solution, (6) a negative control reaction solution and (7) a sample treatment solution. The kit combines simple treatment of samples, RPA and LFA technologies, no simultaneous equipment is required, and all that is required for detection is to boil the samples, so that the kit is very suitable tobe used on site or in fields, can detect and diagnose brucella in veterinarian clinics or farms, has great clinical application value for diagnosis and control of the diseases and has high detectionprecision and broad market prospects.

Description

technical field [0001] The invention belongs to the technical field of medicine and biotechnology, and in particular relates to an RPA-LFD detection kit for detecting Brucella and a use method thereof. Background technique [0002] Brucellosis, also known as Mediterranean relaxation fever, Malta fever, wave fever or undulating fever, is a zoonotic systemic infectious disease caused by Brucella, which can infect animals, such as cattle, sheep, pigs, dogs and camels, Deer and other animals. Its clinical features are long-term fever, hyperhidrosis, arthralgia, hepatosplenomegaly, etc. The chronic phase of the disease may cause damage to multiple organs and systems. [0003] Brucella is a Gram-negative non-motile bacterium, non-capsulated (smooth type with microcapsule), positive for catalase and oxidase, absolutely aerobic, capable of reducing nitrate, intracellular Parasitic, can survive in many kinds of domestic animals. There are six species of Brucella: Brucella malta (B...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/6804C12Q1/04C12R1/01
CPCC12Q1/6804C12Q1/6844C12Q1/689C12Q2521/507C12Q2522/101C12Q2565/625
Inventor 陈泽良刘宝山韩小虎隋昀原张欢刘金玲沈国顺
Owner SHENYANG AGRI UNIV
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