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Kit and method for simultaneous detection of Staphylococcus aureus gene and Escherichia coli gene by using dual molecular beacon-LAMP process

A technology of molecular beacons and staphylococci, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of false positives, aerosol pollution, etc., and achieve the effect of simple operation, strong specificity, and high sensitivity

Active Publication Date: 2013-12-11
SHAANXI UNIV OF SCI & TECH
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Problems solved by technology

However, LAMP products are susceptible to aerosol contamination, causing false positive results

Method used

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  • Kit and method for simultaneous detection of Staphylococcus aureus gene and Escherichia coli gene by using dual molecular beacon-LAMP process
  • Kit and method for simultaneous detection of Staphylococcus aureus gene and Escherichia coli gene by using dual molecular beacon-LAMP process
  • Kit and method for simultaneous detection of Staphylococcus aureus gene and Escherichia coli gene by using dual molecular beacon-LAMP process

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Embodiment Construction

[0047] The present invention will be further described in detail below in conjunction with specific drawings and specific embodiments, which are explanations rather than limitations of the present invention.

[0048] The present invention takes Staphylococcus aureus and Escherichia coli O157:H7 as examples to analyze the detection method of the dual molecular beacon-LAMP, so as to realize rapid and accurate simultaneous detection of two pathogenic bacteria.

[0049]According to the conserved sequences of toxin genes or invasion genes of food-borne pathogens published by GenBank, we selected the thermostable nucase Nuc gene of Staphylococcus aureus and the hemolysin gene rbfE of Escherichia coli O157:H7, and designed LAMP primers and Molecular beacon probes, labeled with different fluorescent agents, established a dual molecular beacon-LAMP method to detect the reaction system of these two foodborne pathogens, and applied this detection system to the detection of common bacteria...

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Abstract

The invention discloses a kit and a method for simultaneous detection of a Staphylococcus aureus gene and an Escherichia coli gene by using a dual molecular beacon-LAMP process, belonging to the technical field of molecular biological detection. The kit comprises an LAMP reaction system, wherein the LAMP reaction system comprises a primer group used for detecting the Staphylococcus aureus gene nuc and an rfbE gene of the Escherichia coli O157: H7, the primer group includes a pair of outer primers F3 and B3, a pair of inner primers FIP and BIP and a molecular beacon probe of the nuc gene and further includes a pair of outer primers F3 and B3, a pair of inner primers FIP and BIP and a molecular beacon probe of the rfbE gene. The method provided by the invention has the advantages of high sensitivity, strong specificity, avoidance of cross reaction with a dozen of bacteria of a control group, simple operation, realization of visual and clear observation of results and a fast detection speed.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and relates to a method for simultaneously detecting two kinds of food-borne pathogenic bacteria by a double molecular beacon-loop-mediated isothermal amplification technology, in particular to a double molecular beacon-LAMP method for simultaneous detection of gold A kit and method for the nuc gene of Staphylococcus aureus and the rfbE gene of Escherichia coli O157:H7. Background technique [0002] Food is food to safety first. The problem of food safety has become a major safety hazard that plagues the world, especially food contamination and food-borne diseases caused by pathogenic microorganisms have become a worldwide public health problem. The World Health Organization (WHO) reports that billions of cases of food-borne diseases occur every year in the world. Diarrhea caused by microbial contamination and death. Commonly important pathogenic bacteria in microbial food ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/14C12Q1/10
Inventor 龚频王兰张涛涛陈福欣
Owner SHAANXI UNIV OF SCI & TECH
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