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Primer for loop-mediated isothermal amplification for Brucella and detection reaction system

A ring-mediated isothermal and Brucella technology, which is applied in the direction of microorganism-based methods, microorganism measurement/inspection, microorganisms, etc., can solve the problems of long time-consuming, difficult grass-roots production, popularization and application, etc., and achieve simple, fast and accurate operation The effect of low requirements for identification and experimental equipment

Inactive Publication Date: 2013-08-14
天津市动物疫病预防控制中心
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Problems solved by technology

At present, the routine diagnostic techniques for Brucella include bacterial isolation and culture, serum agglutination test, complement fixation test, enzyme-linked immunosorbent assay, PCR and other methods. It is difficult to popularize and apply in grassroots production, so it is necessary to establish a simple, fast and accurate pathogenic diagnosis method

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  • Primer for loop-mediated isothermal amplification for Brucella and detection reaction system
  • Primer for loop-mediated isothermal amplification for Brucella and detection reaction system
  • Primer for loop-mediated isothermal amplification for Brucella and detection reaction system

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Embodiment Construction

[0029] Below in conjunction with accompanying drawing and specific embodiment the present invention is described in further detail:

[0030] The primers for Brucella ring-mediated isothermal amplification of the present invention include forward internal primer FIP, forward external primer F3, reverse internal primer BIP, reverse Outward primer B3, the sequence is as follows:

[0031] Outer primer: F3 5'-ATTGCCGGTTCGCAGATC-3'

[0032] B3 5'-GCGGAAATCCTGCGTGTC-3'

[0033] Internal primer: FIP 5'-TCAGCTTGGCTTCGAGACCGCTTAACAACGGCTTGGACGA-3'

[0034] BIP 5'-GGCCGCGTTGAGTACCGTTAAGCTTGTTGCGAACAGTCG-3'

[0035] The primer is complementary to a part of the nucleic acid sequence at position 412-618bp of the gene omp25 or its complementary chain.

[0036] Using the above-mentioned detection reaction system for the primers used for Brucella loop-mediated isothermal amplification, the 25 μL reaction system is:

[0037] Outer primer F3: 5pmol·L -1 , 0.5μL

[0038] Outer primer B3: 5...

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Abstract

The invention discloses a primer for loop-mediated isothermal amplification for Brucella and a detection reaction system. Four LAMP specific primers are designed for a conservative OMP25 protein gene of Brucella, and an LAMP quick detection method of Brucella is established by optimizing the reaction condition. Specificity and sensitivity test results show that the method can specifically detect Brucella, wherein the minimum limit of detection is 5 copies / microlitre. The amplification product can be judged through agarose gel electrophoresis and chromogenic reaction. LAMP can quickly, intuitively and accurately detect Brucella, and is a detection method suitable for field application of basic level.

Description

technical field [0001] The invention relates to a primer and a reaction detection system for isothermal amplification mediated by Brucella rings. Background technique [0002] Brucellosis is a zoonotic disease characterized by abortion and fever caused by Brucella, which seriously threatens the life and health of humans and various animals, causing huge economic losses and serious public health hazards . The key to the prevention, control and treatment of brucellosis lies in the rapid, accurate and early detection and diagnosis of pathogenic microorganisms. At present, the routine diagnostic techniques for Brucella include bacterial isolation and culture, serum agglutination test, complement fixation test, enzyme-linked immunosorbent assay, PCR and other methods. It is difficult to popularize and apply it in grass-roots production, so it is necessary to establish a simple, fast and accurate pathogenic diagnosis method. [0003] Notomi et al. invented the loop-mediated iso...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12Q1/68C12N15/11C12R1/01
Inventor 李秀梅
Owner 天津市动物疫病预防控制中心
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