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Multiplex PCR (polymerase chain reaction) method for testing three main pathogens in beef

A detection method and technology for pathogenic bacteria, applied in the biological field, can solve the problems of long detection period of pathogenic bacteria, cumbersome operation, low specificity, etc., and achieve the effects of saving expenses, reducing operating steps, and high specificity

Inactive Publication Date: 2014-12-24
SHANDONG AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a multiple PCR detection method for three main pathogenic bacteria in beef, aiming to solve the problems of long detection period, cumbersome operation and low specificity of traditional food-borne pathogenic bacteria

Method used

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  • Multiplex PCR (polymerase chain reaction) method for testing three main pathogens in beef
  • Multiplex PCR (polymerase chain reaction) method for testing three main pathogens in beef
  • Multiplex PCR (polymerase chain reaction) method for testing three main pathogens in beef

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Embodiment Construction

[0036] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0037] figure 1 Shown in beef of the present invention three kinds of main pathogenic bacteria multiple PCR detection method flow process, as shown in the figure, the present invention is realized in this way, three kinds of main pathogenic bacteria multiple PCR detection methods in a kind of beef comprise:

[0038] S101, strain culture:

[0039] Under sterile conditions, pick appropriate amount of Salmonella, Listeria monocytogenes, and Escherichia coli O157:H7 with an inoculation loop and inoculate them in TSB, culture with shaking at 37°C for 18 hours, count Salmonella on HE agar plates, and u...

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Abstract

The invention discloses a multiplex PCR method for testing three main pathogens in beef. The method includes the steps of culturing bacterial strains of salmonella, listeria monocytogenes, escherichia coli O157:H7; contrasting propagation effects of different propagation broth on three pathogens; selecting primers; amplifying through a multiplex PCR system; extracting DNA (Deoxyribose Nucleic Acid) of bacteria in a beef sample to be tested; building multiplex PCR system parameters. By means of the multiplex PCR method for testing three main pathogens in beef, high specificity and sensitivity of conventional PCR are maintained, operation steps are reduced, and the purpose of one-time amplification for simultaneously testing a variety of microorganisms is achieved, so that time and costs are greatly saved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a multiplex PCR detection method for three main pathogenic bacteria in beef. Background technique [0002] Food safety has become a public health issue of common concern all over the world. Among various food safety incidents, foodborne diseases caused by bacteria are the most and involve the most extensive number of people. Salmonella, Listeria monocytogenes, and E. coli O157:H7 are considered to be the three most important foodborne pathogens in beef. In China, 70%-80% of bacterial food poisoning is caused by Salmonella, and most of them come from animal-derived food, and Salmonella food poisoning caused by beef accounts for more than 40%. Listeria monocytogenes can cause listeriosis, and the mortality rate is as high as 20%-30%, especially for pregnant women, children, the elderly and people with low immunity. Raw meat products such as beef, pork, lamb and poultry a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/19C12R1/42C12R1/01
CPCC12Q1/686C12Q2537/143
Inventor 罗欣朱立贤宋东晓毛衍伟张一敏梁荣蓉牛乐宝
Owner SHANDONG AGRICULTURAL UNIVERSITY
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