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Legionnella rapid detecting and parting method

A typing method, Legionella technology, applied in the field of rapid detection and typing of Legionella, can solve the problems of long time, complexity, and low specificity, and achieve the effect of simple detection

Active Publication Date: 2010-06-02
重庆金域医学检验所有限公司
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Problems solved by technology

[0007] The purpose of the present invention is to provide a kind of method that can identify Legionella and rapidly molecularly type it into Legionella pneumophila for the above-mentioned legionella molecular typing method. A rapid, simple, specific and sensitive rapid detection and typing method for Legionella and non-pneumophila Legionella

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  • Legionnella rapid detecting and parting method

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Embodiment Construction

[0025] Principle Description

[0026] The present invention utilizes the high conservation of a 226bp base sequence in the Legionella 16S rRNA gene in Legionella, can design a pair of primers to amplify the base fragment, and use PCR to identify Legionella and non-Legionella, namely Legionella DNA as template DNA can amplify the target band under certain reaction conditions and reaction system, while non-Legionella genomic DNA as template cannot amplify the target band under the reaction conditions and reaction system . According to the bioinformatics analysis of the band for this purpose, such as figure 1 As shown, by analyzing the sequence of the target band known in the gene bank, it can be found that some special base sites "ACNGT" are in the target band obtained by PCR using Legionella pneumophila genomic DNA as a template Both exist in the specific 178-182 position in the band, while using the non-Legionella pneumophila genomic DNA as a template, the target band obtain...

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Abstract

The invention discloses a legionnella rapid detecting and parting method, comprising the steps of: (1) extracting a genom DNA of a detecting sample, (2) performing PCR by using genom DNA as a template in order to acquire PCR products, (3) performing agarose gel electrophoresis analysis to the PCR products in order to determine if legionnella exists, (4) performing digestion reaction to the detected positive PCR products and then performing agarose gel electrophoresis analysis to the digestion reaction products and parting molecule. Compared with the prior art, the legionnella rapid detecting and parting method of the invention can rapidly and simply detect the legionnella and can part the molecule thereof into legionella pneumophilia and non-pneumophilia legionella.

Description

Technical field: [0001] The invention relates to the technical field of Legionella detection and typing methods, in particular to a rapid detection and typing of Legionella that can not only detect Legionella, but also can type Legionella into Legionella pneumophila and non-pneumophila Legionella method. Background technique: [0002] Modern molecular typing techniques mainly include plasmid DNA map analysis, chromosomal DNA analysis after restriction endonuclease digestion, nucleic acid probe hybridization, pulsed field gel electrophoresis (PFGE), PCR, and multilocus sequence analysis (MLST) techniques, etc. . DNA repeat sequence PCR (Rep-PCR) has strong practicability, low cost and good reproducibility, and can be used for census work. PFGE technology has high resolution and is the best choice for molecular typing of pathogenic microorganisms. The new MLST technology with higher resolution and reproducibility is gradually being used. But overall, analysis of chromosoma...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12R1/01
Inventor 詹晓勇朱庆义胡朝晖李连青张远志
Owner 重庆金域医学检验所有限公司
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