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Primer and probe sequence for multifluorescent PCR synchronous detection of Salmonella, Vibrio parahaemolyticus and Escherichia coli O157:H7

A technology of Escherichia coli and O157, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, and resistance to vector-borne diseases, etc., can solve the problems of economic loss, complicated operation, and retention of enterprises, and achieve convenient detection and use and save detection reagents , the effect of reducing the cost of testing

Inactive Publication Date: 2009-05-27
中华人民共和国汕头出入境检验检疫局
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AI Technical Summary

Problems solved by technology

However, there is no method and related standards that can simultaneously detect multiple pathogenic bacteria in aquatic products
Although there are various rapid detection methods for the detection of these three pathogenic bacteria, such as immunomagnetic strain enrichment method, enzyme-linked immunosorbent assay, automatic enzyme-linked immunofluorescence method, chromogenic medium method, DNA probe technology, PCR technology, etc., but these methods have not been widely promoted due to their shortcomings of one kind or another. In fact, the inspection of pathogenic bacteria in aquatic products by relevant departments in my country basically remains on the traditional bacterial isolation and cultivation, which is not only time-consuming, but often requires 4 ~7 days, it may also be missed due to complicated operations and many reagents used
From January to July 2005, 11 batches of my country's aquatic products exported to the United States were detained due to the detection of Salmonella by the US FDA, causing large economic losses to the enterprise

Method used

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  • Primer and probe sequence for multifluorescent PCR synchronous detection of Salmonella, Vibrio parahaemolyticus and Escherichia coli O157:H7
  • Primer and probe sequence for multifluorescent PCR synchronous detection of Salmonella, Vibrio parahaemolyticus and Escherichia coli O157:H7
  • Primer and probe sequence for multifluorescent PCR synchronous detection of Salmonella, Vibrio parahaemolyticus and Escherichia coli O157:H7

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Embodiment

[0048] Detection kit: pathogenic bacteria in aquatic products (Salmonella, Vibrio parahaemolyticus and Escherichia coli O 157 :H 7 ) multiple fluorescent PCR detection kit, which can be used for detection of 20 aquatic samples. The specific composition is as follows:

[0049] Composition (20 samples) Specification DNA extraction solution 18mL 1 bottle PCR reaction solution. 240μL 1 tube primer probe 180μL 1 tube positive control template 20μL 1 tube negative control template 20μL 1 tube Deionized water 1mL 1 tube

[0050] Sample 1: Artificially added appropriate concentrations of Salmonella, Vibrio parahaemolyticus and Escherichia coli O 157 :H 7 Shrimp with three pathogenic bacteria.

[0051] Sample 2: No Salmonella, Vibrio parahaemolyticus and Escherichia coli O were confirmed by SN standard test 157 :H 7 Shrimp with three pathogenic bacteria.

[0052] Detection steps: use the above-mentioned kits and samples f...

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Abstract

The invention provides a primer and a probe sequence for multiple fluorescence PCR synchronous detection of nucleotide segments of three types of malignant bacteria of salmonella, vibrio parahaemolyticus and bacillus coli O157:H7. The primer and the sequence are as shown in figures.

Description

【Technical field】 [0001] The present invention relates to a method for Salmonella, Vibrio parahaemolyticus and Escherichia coli O 157 :H 7 Primer and probe sequences for multiplex fluorescent PCR simultaneous detection of nucleotide fragments of three pathogenic bacteria. 【Background technique】 [0002] Salmonella, Vibrio parahaemolyticus, and Escherichia coli O 157 :H 7 It is an important food-borne pathogen. It mainly infects humans through contaminated animal-derived food such as aquatic products, causing food poisoning, typhoid fever, uremia, etc., and can lead to death in severe cases. Therefore, these three pathogenic bacteria have attracted much attention, and they are pathogenic bacteria that must be inspected or monitored for imported and exported aquatic products and other animal-derived products in my country and most developed countries in the world. [0003] Aquatic products are the main agricultural products exported to Europe and the United States, and hav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/10C12Q1/68C12N15/11
CPCY02A50/30
Inventor 许如苏林志雄林彩华蔡颖陈其生陈冠武
Owner 中华人民共和国汕头出入境检验检疫局
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