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Salmonella constant temperature gene amplification fast detecting kit and method

A technology of constant temperature gene amplification and detection kits, which is applied in biochemical equipment and methods, measurement/inspection of microorganisms, and material analysis through observation of the influence of chemical indicators, etc., which can solve harsh, limited development, and unclear Accurate and other issues, to achieve the effect of high yield, simple identification, high sensitivity

Inactive Publication Date: 2008-08-20
SOUTH CHINA UNIV OF TECH
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AI Technical Summary

Problems solved by technology

Although the disclosed LAMP technology has high sensitivity and can efficiently amplify nucleic acid under isothermal conditions, the existing technology also has shortcomings, mainly: 1. Its special primer requirements are extremely harsh and limit the wide application of this method; 2. .It is not clear and accurate enough to judge the result only by the milky white precipitate generated by the reaction, and it is not easy to realize the miniaturization of the reaction, which limits its further development

Method used

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Embodiment Construction

[0024] Prepare the Salmonella isothermal gene amplification rapid detection kit according to the following formula:

[0025] ——Primer mixture: the concentration of the four primers is 10 pmol / μl, and the primer sequences are such as SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, and SEQ ID NO.4;

[0026] ——Bst DNA polymerase: the concentration is 8 U / μl;

[0027] ——Reaction solution: 80μl 10mM dNTP, 50μl 10×ThermoPol reaction buffer, 20μl 150mM MgSO 4 .

[0028] - Sample pretreatment solution: 20mM Tris-HCl (pH 8.0), 2mM EDTA, 1.2% TritonX-100.

[0029] —— Chromogenic agent: fluorescent dye 1×SYBR Green I.

[0030] ——The positive control is the genomic DNA of Salmonella choleraesuis, and the negative control is the reaction mixture without template.

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Abstract

The present invention discloses a salmonella constant-temperature gene amplification fast detecting reagent kit and the method thereof, the reagent kit comprises the primer mixed liquid, the Bst DNA polyase, the reacting liquid, the sample pretreating liquid, the color developing agent, the positive comparison and negative comparison, wherein the sequence of the primer is the sequence such as SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4. Through extracting the salmonella genome DNA, constant-temperature gene amplification reaction and judging the reaction product result with the reagent kit the high-specificity, quick, high-sensitivity and simple detecting to the salmonella is realized.

Description

technical field [0001] The invention relates to a microorganism detection reagent and method, in particular to a rapid detection kit and method for constant temperature gene amplification of Salmonella. Background technique [0002] Salmonella is a major genus of food-borne pathogens. Among various bacterial food poisoning incidents, food poisoning caused by Salmonella ranks at the forefront. It is mainly transmitted through food, especially animal food, such as eggs, milk and meat products, causing human gastrointestinal diseases. Salmonella is increasingly threatening human health. Therefore, it is necessary to establish a safe, reliable, simple and rapid detection method to ensure food safety. Conventional Salmonella detection methods include bacterial proliferation and culture, biochemical identification, and serological identification. It usually takes 4 to 6 days to complete, which is not only time-consuming and laborious, but can no longer meet the control requiremen...

Claims

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Application Information

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IPC IPC(8): G01N21/78C12Q1/68C12Q1/24
CPCY02A50/30
Inventor 顿博影李琳石磊王丽耿予欢
Owner SOUTH CHINA UNIV OF TECH
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