Listeria monocytogenes nucleic acid chromatography detection kit, its detection method and application thereof

A technology for detecting kits and bacterial nucleic acids, which is applied in the fields of molecular biology and immunology, can solve the problems of long detection time and cumbersome steps, and achieve the effects of low detection cost, simple operation process, and reduced detection cost

Inactive Publication Date: 2012-06-27
北京陆桥技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem solved by the present invention is to provide a nucleic acid chromatography kit for the detection of Listeria monocytogenes in food and its detection method and application; the main pu

Method used

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  • Listeria monocytogenes nucleic acid chromatography detection kit, its detection method and application thereof
  • Listeria monocytogenes nucleic acid chromatography detection kit, its detection method and application thereof
  • Listeria monocytogenes nucleic acid chromatography detection kit, its detection method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0063] The detection specificity of embodiment 1 kit

[0064] Listeria ovis (1), Listeria innocua (2), Listeria sierra (3), Listeria weideri (4), Listeria gasseri (5) It is a verified strain within the genus of Listeria, other common pathogenic bacteria Escherichia coli (6), Salmonella enteritidis (7), Bacillus subtilis (8), Staphylococcus aureus (9), Shigella flexneri Bacteria (10) was used as a verified bacterial strain outside the genus of Listeria, and its genome was extracted as a template for PCR amplification, and Listeria monocytogenes was used as a positive control C + , see the test results figure 2 .

[0065] It can be seen from the results in the figure that only in the positive control (C + ) shows positive, and the rest are negative, indicating that the kit of the present invention has good specificity and accurate detection results.

Embodiment 2

[0066] The sensitivity of embodiment 2 kits

[0067] Based on the DNA of Listeria monocytogenes, make a gradient dilution concentration ( / μL) of 1ng, 100pg, 10pg, 1pg, and 100fg respectively as a template for PCR amplification, and detect it with a test strip, and set a negative control at the same time (C - ), see the result image 3 .

[0068] As can be seen from the results, it is weakly positive at 1pg, and the detection line at 100fg is negative if it does not develop color, indicating that the sensitivity of the kit of the present invention to detect Listeria monocytogenes can reach 1pg.

Embodiment 3

[0069] Example 3 Sample Detection

[0070] A total of 50 samples of dairy products, eggs, vegetables, fish, shrimp, chicken, pork, etc. were purchased from supermarkets and markets. After enrichment, the enrichment solution was taken to extract the genome for PCR amplification. The products were tested with test strips. The results are shown in In Table 2, there is no significant difference compared with the results of the traditional microbial method GB 4789.30-2010, and the coincidence rate is 97.7%.

[0071] Table 2 Sample detection test

[0072]

[0073] χ2=0 P>0.03

[0074] The above results show that the kit of the present invention has a high coincidence rate compared with the traditional microbial method, saves time and has no missing detection phenomenon. The traditional method has certain requirements on the effect of sample enrichment. After the positive sample is enriched to a certain concentration, the suspicious colony can be displayed by streaking on the pl...

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Abstract

The invention discloses a listeria monocytogenes nucleic acid chromatography detection kit, its detection method and an application thereof, and belongs to the field of molecular biology and immunology. According to the invention, with the combination of a polymerase chain reaction technology in molecular biology and an immunochromatography technology, rapid detection of listeria monocytogenes in food is accomplished. The kit provided by the invention includes three parts of DNA extraction, PCR amplification and strip detection. The combination of the molecular biology technology and the immunochromatography technology is accomplished by modification of primers and selection of corresponding coatings and markers in strip preparation so as to achieve specific amplification product detection of listeria monocytogenes in food. Compared with a traditional biochemistry method, the method provided by the invention is used to greatly save detection time and simplify operation steps, has characteristics of accuracy, rapidity, convenience and safety, and is suitable for detection of a large number of samples.

Description

technical field [0001] The invention belongs to the fields of molecular biology and immunology, and relates to a kit and a method for rapidly detecting Listeria monocytogenes in food. Background technique [0002] As the country pays more and more attention to food safety and import and export trade, the requirements for food safety testing technology are also getting higher and higher. According to the requirements for the development of food safety and entry-exit inspection and quarantine work in the field of public security in the "National Medium and Long-Term Science and Technology Development Plan (2006-2020)", research on efficient, fast and accurate detection technology has become a topic in this field. One of the main trends and goals. [0003] According to the information held by the World Health Organization, among the risk factors of foodborne diseases, microbial food poisoning is still the primary hazard; in my country, according to the statistics of the Chines...

Claims

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Application Information

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IPC IPC(8): G01N33/569
Inventor 王丽丽赵瑜唐慧林何艳玲李瑾
Owner 北京陆桥技术股份有限公司
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