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Novel colorimetric sensing method for dual pathogenic bacteria

A detection method and nucleotide sequence technology, applied in the field of biological detection, can solve the problems of difficult analysis results, cumbersome experimental operations, cumbersome operations, etc., and achieve the effect of simple and efficient detection and analysis

Inactive Publication Date: 2018-07-06
CHINA AGRI UNIV
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Problems solved by technology

[0002] The traditional bacterial detection method is mainly based on the physiological and biochemical characteristics of the bacteria. After the steps of pre-enrichment, selective plate separation, and biochemical identification, it takes 5-7 days from sampling to confirming the results. The detection cycle is long, the operation is cumbersome, and the workload is heavy.
Using the specificity of antigen-antibody reaction to identify bacteria has a history of more than half a century, but the screening of microbial antibodies is very cumbersome, and the final detection specificity is not high
The continuous improvement and development of molecular biology detection technology has overcome the problems of cumbersome and time-consuming experimental operations of traditional detection methods, and has also led to the rapid development of rapid detection methods for microorganisms. However, the disadvantage of molecular biology methods is that the results are not visualized. Not easy to analyze results

Method used

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  • Novel colorimetric sensing method for dual pathogenic bacteria
  • Novel colorimetric sensing method for dual pathogenic bacteria
  • Novel colorimetric sensing method for dual pathogenic bacteria

Examples

Experimental program
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Effect test

Embodiment 1

[0119] Example 1. Establishment of a new ultra-fast PCR-based dual colorimetric sensing method for the detection of Salmonella and Staphylococcus aureus

[0120] (1) Experimental materials

[0121] The strain information used in this example is shown in Table 1, and the nucleotide sequences of the designed primers are shown in Table 2 and the sequence list.

[0122] Table 1

[0123]

[0124] Table 2

[0125]

[0126] In Table 2, the nucleotide sequence on the left side of the connecting arm (oxyethyleneglycol bridge) of the upstream primer Primer 1 is the nucleotide sequence shown in SEQ ID No. 1 in the sequence table, and the nucleotide sequence on the right side of the connecting arm is the sequence The nucleotide sequence shown in SEQID №: 2 in the list, the chemical structure of the connecting arm is:

[0127]

[0128] In Table 2, the nucleotide sequence of the downstream primer Primer 2 is the nucleotide sequence shown in SEQ ID No.: 3 in the sequence listing....

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Abstract

The invention discloses a novel colorimetric sensing method for dual pathogenic bacteria, and the novel colorimetric sensing method is used for supersensitive detection of salmonella and staphylococcus aureus. The method establishes an ultra-quick PCR reaction system, so that a conventional PCR process with consumed time of about three hours is reduced to 5 minutes, and therefore, consumed time ofPCR reaction is remarkably reduced; the ultra-quick PCR reaction system loads a nucleic acid self-assembly developing module, so that a reaction signal is amplified again, supersensitive detection ofpathogenic bacteria is favorably realized, and the problem that the conventional PCR results are difficult for visual detection is solved. Compared with a conventional method, the detecting method and a biosensor are quicker, are more sensitive, are stronger in specificity, and are high in sensitivity, and detected results are reliable, so that analyzing and detecting steps can be simplified, analysis time is shortened, and above all, online real-time detection is possible, therefore, carrying and field operating are convenient in the microbiological detection field of the food safety and quick detecting field, and the application prospect is very good.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a new method for colorimetric sensing of dual pathogenic bacteria. Background technique [0002] The traditional bacterial detection method is mainly based on the physiological and biochemical characteristics of the bacteria. After the steps of pre-enrichment, selective plate separation, and biochemical identification, it takes 5-7 days from sampling to confirming the results. The detection cycle is long, the operation is cumbersome, and the workload is heavy. . Using the specificity of antigen-antibody reaction to identify bacteria has a history of more than half a century, but the screening of microbial antibodies is very cumbersome, and the final detection specificity is not high. The continuous improvement and development of molecular biology detection technology has overcome the problems of cumbersome and time-consuming experimental operations of tr...

Claims

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Application Information

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IPC IPC(8): C12Q1/686C12Q1/689C12Q1/14C12Q1/10C12R1/445C12R1/42
CPCC12Q1/686C12Q1/689C12Q2600/16C12Q2537/143C12Q2525/301C12Q2563/125C12Q1/6853C12Q2525/186C12Q2525/197
Inventor 罗云波许文涛黄昆仑田晶晶杜再慧
Owner CHINA AGRI UNIV
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