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Non-diagnostic method for detecting coxiella burnetii with loop-mediated isothermal amplification (LAMP) technology

A technology based on Coxa Bainiette and ring-mediated isothermal technology, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection, can solve the problem of low signal-to-noise ratio of results and achieve detection cost Low cost, simple operation, easy operation effect

Active Publication Date: 2013-01-02
CHINESE ACAD OF INSPECTION & QUARANTINE
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Problems solved by technology

[0004] Aiming at the problems in the prior art that the results can only be observed through electrophoresis gel imaging, or the signal-to-noise ratio of the results displayed by the turbidimeter is low, the purpose of the present invention is to provide a device with the advantages of simple operation, fast response, and short time consumption. The Loop-Mediated Isothermal Amplification Technique as a Non-Diagnostic Approach for the Detection of Coxiella beinii

Method used

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  • Non-diagnostic method for detecting coxiella burnetii with loop-mediated isothermal amplification (LAMP) technology
  • Non-diagnostic method for detecting coxiella burnetii with loop-mediated isothermal amplification (LAMP) technology
  • Non-diagnostic method for detecting coxiella burnetii with loop-mediated isothermal amplification (LAMP) technology

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Experimental program
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Embodiment

[0075] Comparison of Blood Sample Tests

[0076] (1) Take 30 blood samples, and use the genomic DNA extraction kit to extract DNA;

[0077] (2) Take a part of the extracted DNA solution for ordinary PCR amplification;

[0078] (3) The remaining DNA solution was subjected to LAMP amplification of Coxiella basilaris, using the outer primers F3, B3 and inner primers FIP, BIP, extracted genomic DNA, enzyme, and buffer to prepare a 25 μL reaction system;

[0079] (4) Loop-mediated isothermal amplification procedure;

[0080] (5) The results of electrophoresis detection and analysis were compared with the PCR amplification of Coxiella commonis and the LAMP amplification of Coxiella bezii.

[0081] As a result, 2 samples were positive, which was consistent with the results obtained by ordinary PCR, indicating that the established method was not different from the ordinary PCR method.

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Abstract

The invention discloses a non-diagnostic method for detecting coxiella burnetii with a loop-mediated isothermal amplification (LAMP) technology. An LAMP method has the advantages of easiness in operating, quick reaction, completion within one hour under a constant-temperature condition, low time consumption, one-step completion of reactions and product detection, easiness and convenience in operating and low detection cost. Due to the adoption of the non-diagnostic method, a novel developing direction is provided for the detection of the coxiella burnetii; and the method is expected to become a simple conventional detection means, is particularly suitable for basic level inspection and quarantine institutes, and plays important roles in raising the sanitary level, guaranteeing the human life safety and prompting the development of the sanitary inspection institutions.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a new improved ring-mediated isothermal amplification method and its application in detecting Coxiella bezieri. Background technique [0002] Coxiella burnetti, also known as Coxiella burnetti, commonly known as Q fever Rickettsia, is the pathogen that causes Q fever and belongs to Rickettsiaceae, Rickettsia family, Kirkettsia Steinia, Gram-negative small bacilli or cocci, can pass through bacterial filters. According to electron microscope observation, the whole cell of Cox body of Bernard can be divided into three parts: the outer layer composed of microcapsule, cell wall and cytoplasmic membrane; the dense peripheral layer containing ribosome particles in the cytoplasmic membrane; Central dense body formed by winding filaments. Phylogenetic studies have shown that Coxiella beinii is far from other Rickettsia phylogenetic relationships, it belongs to the γ subg...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/01
Inventor 王静杨宇杨永莉刘丽娟赵婷婷王旺王莎莎
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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