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Detection method for staphylococcus aureus

A staphylococcus and detection method technology, applied in the directions of microorganism-based methods, biochemical equipment and methods, and microbial determination/inspection, can solve the problems of low detection sensitivity and long detection time of Staphylococcus aureus, and reduce detection Lower limit, increased fluorescence intensity, and improved detection efficiency

Inactive Publication Date: 2017-08-08
CHANGSHA MEDICAL UNIV
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Problems solved by technology

[0004] The purpose of this application is to provide a detection method for staphylococcus aureus, to solve the problem of low detection sensitivity and long detection time of staphylococcus aureus

Method used

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  • Detection method for staphylococcus aureus
  • Detection method for staphylococcus aureus
  • Detection method for staphylococcus aureus

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Embodiment Construction

[0039] refer to figure 1 , the embodiment of the present application provides a detection method for Staphylococcus aureus, comprising the following steps:

[0040] Step S1, combining the first probe, the second probe and single-stranded DNA (ssDNA) into a double fluorescently labeled probe (TFP) with two carboxyfluorescein (FAM);

[0041] The DNA sequence of the first probe is 5'-FAM-TCTAGA-3';

[0042] The DNA sequence of the second probe is 5'-TCTAGA-FAM-3';

[0043] The sequence of the single-stranded DNA is 5'-TCTAGAAGTTATCCCAGTCTTATAGGTAGGTTCATGA-3';

[0044] Said forming the first probe, the second probe and the single-stranded DNA into a double fluorescently labeled probe with two carboxyfluoresceins includes:

[0045] Denature 50 nM of the first probe, second probe and single-stranded DNA in a water bath at 95°C for 5 minutes, and then ice-bath for 10 minutes; Mix and incubate at room temperature for 30 min to form a double fluorescently labeled probe with two car...

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Abstract

The invention discloses a detection method for staphylococcus aureus. The detection method comprises the following steps: preparing a double-fluorescence-label probe with two carboxy flouorescein labels from a first probe, a second probe and a single-chain DNA; mixing the double-fluorescence-label probe with a graphene oxide solution, thus forming a compound solution; extracting the total RNA of a bacteria solution to be detected; simultaneously adding the total RNA and deoxyribonuclease into the compound solution, thus forming a mixed solution, and incubating the mixed solution; measuring a fluorescence value of the incubated mixed solution, and calculating the concentration of the staphylococcus aureus according to the fluorescence value. According to the detection method disclosed by the embodiment of the invention, a section of specifically recognized staphylococcus aureus 16S rRNA is used as a target to design the probe, and a double-fluorescent-label probe / graphene oxide system is built based on the fluorescence superposition principle; cyclic detection on the target is realized according to the characteristic of the deoxyribonuclease, so that a fluorescence signal is amplified, the sensitivity of the detection method for the staphylococcus aureus is improved, and the detection efficiency of the detection method is also improved.

Description

technical field [0001] The present application relates to the technical field of detection of pathogenic bacteria, in particular to a method for rapid detection of Staphylococcus aureus with high sensitivity, which belongs to the interdisciplinary technical field of microbiology and chemistry. Background technique [0002] Staphylococcus aureus is a common pathogenic and lethal pathogen, which can cause a series of diseases such as local suppurative infection, necrotizing pneumonia, pericarditis and food poisoning, and even systemic infection such as sepsis and septicemia. Therefore, rapid and effective detection technology becomes the key to prevention. [0003] With the development of science and technology, various new technologies have been applied to the detection of Staphylococcus aureus, and immunological methods, electrochemical methods, PCR and other methods have been developed, but these methods have the disadvantages of easy contamination, weak selectivity, cumber...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/14C12R1/445
CPCC12Q1/682C12Q1/689C12Q2521/301C12Q2563/107
Inventor 贺气志罗怀青李建明唐亮郝一
Owner CHANGSHA MEDICAL UNIV
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