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Food foodborne pathogenic bacteria rapidly identifying method

A food-borne pathogenic bacteria and identification method technology, which is applied in the field of rapid identification of food-borne pathogenic bacteria, can solve the problems of complicated operation process, time-consuming and laborious, etc., and achieve the effects of high sensitivity, simplified operation procedures and time saving

Inactive Publication Date: 2018-08-17
芜湖市食品药品检验中心
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Traditional food microbial detection usually uses agar plate culture method, which mainly relies on culture medium for culture, separation and biochemical identification. The operation process is complicated and time-consuming, and generally takes several days to complete.

Method used

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Examples

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Embodiment 1

[0035] A rapid identification method for foodborne pathogens in catering food, including the following steps:

[0036] 1) DNA template extraction:

[0037] Take 25 g of food samples, add them to 225 mL of enrichment medium, mix well, and cultivate for 18 hours. Take 50 μL of the bacterial suspension of the standard strain and the bacterial suspension of the sample, add 3 mL of LB broth to 37 ° C, 150 r / min and shake for 8 h, and then extract the template DNA according to the instructions of the bacterial genomic DNA extraction kit.

[0038] 2) Primer screening and reaction system optimization:

[0039] Select the specific genes nuc, invA, prfA of Staphylococcus aureus, Salmonella, and Listeria monocytogenes, and use Primerpremier5.0 and Oligo6.0 software to design 3 pairs of primers, as shown in Table 1 below. Screen the combination suitable for multiplex PCR, react each pair of primers under the same conditions, adjust the amount of primers and the ratio of primer concentra...

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Abstract

The invention discloses a food foodborne pathogenic bacteria rapidly identifying method. The food foodborne pathogenic bacteria rapidly identifying method comprises the steps of 1) DNA (deoxyribonucleic acid) template extraction, 2) primer screening and reaction system optimization including selecting the specific genes of nuc, invA and prfA of staphylococcus aureus, salmonellas and listeria monocytogenes to separately design primer pairs, 3) PCR (polymerase chain reaction) amplification, 4) PCR-DHPLC (polymerase chain reaction-denaturing high performance liquid chromatography) detection of strain DNA, during which only strains coinciding with target amplification can be detected with amplification absorption peaks while other non-targeted amplification bacteria detected to be negative. The food foodborne pathogenic bacteria rapidly identifying method can detect three foodborne pathogenic bacteria in food when applied every single time, thereby saving time and consumable agent, simplifying operation, avoiding excessive artificial errors and achieving high sensitivity.

Description

technical field [0001] The invention belongs to the field of food detection, and particularly relates to a rapid identification method for food-borne pathogenic bacteria in food and beverages. Background technique [0002] In recent years, there have been frequent problems with food safety in our country, and food safety has become an indispensable topic in life. Relevant data from my country Center for Disease Control and Prevention shows that food safety incidents caused by food-borne pathogens are the most serious in my country in recent years. The latest active surveillance of foodborne diseases in China shows that on average, 1 in 6.5 people in my country suffers from foodborne diseases. Mild cases often appear with symptoms of acute gastroenteritis, such as vomiting, nausea, abdominal pain, diarrhea, fever, etc. After treatment, they can recover to health; but severe cases may have respiratory, circulatory, nervous and other system symptoms, and timely rescue can turn...

Claims

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Application Information

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IPC IPC(8): C12Q1/686C12Q1/14C12Q1/04C12R1/445C12R1/42C12R1/01
CPCC12Q1/686C12Q2537/143C12Q2565/137
Inventor 董菁刘丽吴静胡孔兴刘存军张敏敏杨浩毅
Owner 芜湖市食品药品检验中心
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