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A kind of HDA primer of Clostridium perfringens in food and application method thereof

A Clostridium perfringens, food technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problem that nucleic acid amplification technology cannot be more widely promoted and applied, and achieve considerable economic Benefits, saving manpower and material resources, and the effect of rapid detection methods

Active Publication Date: 2016-08-24
河北省食品检验研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, PCR technology has always been unable to get rid of the limitation of relying on sophisticated instruments and equipment, making PCR-based nucleic acid amplification technology unable to be more widely promoted and applied.

Method used

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  • A kind of HDA primer of Clostridium perfringens in food and application method thereof
  • A kind of HDA primer of Clostridium perfringens in food and application method thereof
  • A kind of HDA primer of Clostridium perfringens in food and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Design of C. perfringens HDA primers

[0032] The primers were synthesized by Bao Bioengineering (Dalian) Co., Ltd. The sequences of the two primers are shown in Table 1.

[0033] Table 1 Clostridium perfringens HDA primer sequence list

[0034] Primername

Embodiment 2

[0035] Example 2: Application of primers in LAMP detection of Staphylococcus aureus

[0036] (1) Extraction of genomic DNA from Clostridium perfringens

[0037] Pick out Clostridium perfringens and add it to a conical flask with 1% peptone water, shake it slowly by hand until it is fully mixed, and incubate at 36℃±1℃ for 24h±2h. Pipette 1mL of transfection into 10mL SPS, and incubate at 6℃±1℃ for 24h±2h. Finally, 1 mL of the enrichment solution was taken to extract the genomic DNA of Clostridium perfringens with a bacterial genomic DNA extraction kit, and stored at -20°C for later use.

[0038] (2) Optimization and establishment of HDA reaction system and reaction conditions

[0039] A one-step HDA reaction system was used. The reaction system for detecting Clostridium perfringens by HDA method was: 5 μL of 10×buffer (100 mM dithiothreitol, 350 mM Tris-HAc, 100 mM MgSO) 4 , 1mg / mL BSA), 0.04μmol dNTPs, 0.16μmol ATP, 10U Bst ploymerase, 0.10~0.30μg UvrD helicase, 1.0~5.0μg ...

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PUM

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Abstract

The invention discloses HDA primers of clostridium perfringens in food and application method thereof. The HDA primers include a forward primer shown in SEQ ID NO: 1 and a reverse primer shown in SEQ ID NO: 2. The HAD primers are used for detecting the clostridium perfringens, and the application method of the HAD primers is simple and fast, is simple to operate, is wide in coverage and can be directly used for detection, and can be used for accurately and fast detecting whether the food has clostridium perfringens in the range regulated by hygienic standard, a mass of manpower and materials resources are saved the considerable economic benefit can be obtained, and more importantly, the method has important significance for the security of the import and export food.

Description

technical field [0001] The invention relates to an HDA primer of Clostridium perfringens in food and an application method thereof. Background technique [0002] The main food safety problem in my country is the food-borne diseases caused by the contamination of food by pathogenic microorganisms. Among them, microbial food poisoning ranks first among the factors affecting food safety in my country. Children, pregnant women, the elderly and the infirm, and people with low immunity easier to be victimized. Clostridium perfringens (C.perfringens) is the most common type of Clostridium in clinical gas gangrene, because it can decompose sugar in muscle and connective tissue, produce a lot of gas, cause severe emphysema, and then affect Blood supply, resulting in large-scale necrosis of tissue, and the bacteria can form capsules in the body, hence the name Clostridium capsularis. [0003] The traditional detection method of Clostridium perfringens is cumbersome and time-consuming...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/145
CPCC12Q1/6844C12Q1/689C12Q2521/513
Inventor 周巍张岩张薇王赞章晶晶王爽刘东李强
Owner 河北省食品检验研究院