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Detecting kit for five kinds of food-borne pathogenic bacteria and detection method

A food-borne pathogenic bacteria and detection kit technology, applied in the fields of molecular biology and food safety detection, can solve the problems of unfavorable food quality control, poor detection sensitivity, long operation time, etc., and achieve fast detection speed and low consumption The effect of short time and simple detection procedure

Active Publication Date: 2015-12-23
DALIAN NATIONALITIES UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional detection of pathogenic bacteria uses a series of steps such as bacterial culture, isolation, and identification, which not only takes a long time to operate, but also has poor detection sensitivity, which is not conducive to food quality control

Method used

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  • Detecting kit for five kinds of food-borne pathogenic bacteria and detection method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The test kit includes the following components:

[0049] (1) The concentration of the primer pair for Salmonella is 0.3 μmol / L: upstream primer 5'-GTCCAGCTCTGTCGCCTTAATC-3', downstream primer 5'-ACGGCTCCCTGCTACGCT-3';

[0050] (2) The primer pair concentration of Staphylococcus aureus is 0.3 μmol / L: upstream primer 5'-TCTGATTCAATTCGTGTTTACTATGA-3', downstream primer 5'-CGCTACCTTTAGGCACTGACA-3';

[0051] (3) The primer pair concentration of Escherichia coli O157:H7 is 0.5 μmol / L: upstream primer 5'-TATTTGATGATGATCCAGGGG-3', downstream primer 5'-ATTTTGTTCTCCGTCTTGTCCTA-3';

[0052] (4) The primer pair concentration of Vibrio parahaemolyticus is 0.5 μmol / L: upstream primer 5'-ACTAAGTTGGTCGGTTCGATATG-3', downstream primer 5'-TCTGGCTGCTTAGTTTGTGTTTAG-3';

[0053] (5) The concentration of the primer pair for Listeria monocytogenes is 0.9 μmol / L: upstream primer 5'-GTTAATGAACCTACAAGACCTTCC-3', downstream primer 5'-AACCGTTTCTCCACCATTCCCA-3';

[0054] (6) The concentration of ...

Embodiment 2

[0063] The test kit includes the following components:

[0064] (1) The concentration of the primer pair for Salmonella is 0.5 μmol / L: upstream primer 5'-GTCCAGCTCTGTCGCCTTAATC-3', downstream primer 5'-ACGGCTCCCTGCTACGCT-3';

[0065] (2) The primer pair concentration of Staphylococcus aureus is 0.5 μmol / L: upstream primer 5'-TCTGATTCAATTCGTGTTTACTATGA-3', downstream primer 5'-CGCTACCTTTAGGCACTGACA-3';

[0066] (3) The primer pair concentration of Escherichia coli O157:H7 is 0.3 μmol / L: upstream primer 5'-TATTTGATGATGATCCAGGGG-3', downstream primer 5'-ATTTTGTTCTCCGTCTTGTCCTA-3';

[0067] (4) The primer pair concentration of Vibrio parahaemolyticus is 0.3 μmol / L: upstream primer 5'-ACTAAGTTGGTCGGTTCGATATG-3', downstream primer 5'-TCTGGCTGCTTAGTTTGTGTTTAG-3';

[0068] (5) The concentration of the primer pair for Listeria monocytogenes is 0.7 μmol / L: upstream primer 5'-GTTAATGAACCTACAAGACCTTCC-3', downstream primer 5'-AACCGTTTCTCCACCATTCCCA-3';

[0069] (6) TaqDNA polymerase con...

Embodiment 3

[0078] The test kit includes the following components:

[0079] (1) The concentration of the primer pair for Salmonella is 0.4 μmol / L: upstream primer 5'-GTCCAGCTCTGTCGCCTTAATC-3', downstream primer 5'-ACGGCTCCCTGCTACGCT-3';

[0080] (2) The primer pair concentration of Staphylococcus aureus is 0.4 μmol / L: upstream primer 5'-TCTGATTCAATTCGTGTTTACTATGA-3', downstream primer 5'-CGCTACCTTTAGGCACTGACA-3';

[0081] (3) The primer pair concentration of Escherichia coli O157:H7 is 0.4 μmol / L: upstream primer 5'-TATTTGATGATGATCCAGGGG-3', downstream primer 5'-ATTTTGTTCTCCGTCTTGTCCTA-3';

[0082] (4) The primer pair concentration of Vibrio parahaemolyticus is 0.4 μmol / L: upstream primer 5'-ACTAAGTTGGTCGGTTCGATATG-3', downstream primer 5'-TCTGGCTGCTTAGTTTGTGTTTAG-3';

[0083] (5) The concentration of the primer pair for Listeria monocytogenes is 0.8 μmol / L: upstream primer 5'-GTTAATGAACCTACAAGACCTTCC-3', downstream primer 5'-AACCGTTTCTCCACCATTCCCA-3';

[0084] (6) The concentration of ...

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Abstract

The invention discloses a detecting kit for five kinds of food-borne pathogenic bacteria and a detection method, and belongs to the field of molecular biology and food safety detection. The kit is mainly prepared from primer pairs of salmonella, primer pairs of staphylococcus aureus, primer pairs of escherichia coli O157:H7, primer pairs of vibrio parahaemolyticus, primer pairs of listeria monocytogenes, Taq DNA polymerase, 10*polymerase chain reaction buffer, deoxy-ribonucleoside triphosphate and MgCl2. In the detection process, a to-be-detected sample polymerase chain reaction template is prepared and added to the kit for polymerase chain reaction amplification, and then amplification products are subjected to electrophoresis and positive control. According to the detection method, the five kinds of pathogenic bacteria in food can be detected at the same time, and detection sensitivity is high. The detection process is simple, and popularization is easy.

Description

technical field [0001] The invention relates to the fields of molecular biology and food safety detection, in particular to a detection kit and a detection method for five kinds of food-borne pathogenic bacteria. Background technique [0002] Food safety issues are related to the national economy and people's livelihood. With the continuous development of economy and society, more and more requirements are put forward for the detection of food-borne pathogenic bacteria. Salmonella, Staphylococcus aureus, Escherichia coli O157:H7, Listeria monocytogenes, and Vibrioparahaemolyticus are food-borne pathogens . It is prevalent in general fresh food such as vegetables, meat and poultry, seafood and their processed products, endangering human health and even causing large-scale infectious diseases. The traditional detection of pathogenic bacteria uses a series of steps such as bacterial culture, isolation, and identification, which not only takes a long time, but also has poor de...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/14C12Q1/04C12R1/42C12R1/445C12R1/19C12R1/63C12R1/01
CPCC12Q1/686C12Q1/689C12Q2600/16C12Q2565/125
Inventor 胡文忠姜爱丽何煜波冯可刘星伯杨柳钱红玫李鹤
Owner DALIAN NATIONALITIES UNIVERSITY
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