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Method for identifying foodborne pathogen by surface enhanced Raman spectroscopy

A technology of surface-enhanced Raman for food-borne pathogenic bacteria, applied in biochemical equipment and methods, measurement/inspection of microorganisms, resistance to vector-borne diseases, etc., can solve problems such as time-consuming, high cost, and complicated steps

Inactive Publication Date: 2011-07-06
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although these detection methods are very professional and classic, their steps are complex, time-consuming, and costly, and their main disadvantage is that pre-enrichment must be carried out, and from sampling to determining whether to identify microorganisms qualitatively or quantitatively, the minimum It takes more than one day, so it is difficult to adapt to the needs of the rapid development of modern food production and distribution

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  • Method for identifying foodborne pathogen by surface enhanced Raman spectroscopy
  • Method for identifying foodborne pathogen by surface enhanced Raman spectroscopy
  • Method for identifying foodborne pathogen by surface enhanced Raman spectroscopy

Examples

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preparation example Construction

[0036] (1) Preparation of microbial samples

[0037] Activation of food-borne pathogenic bacteria strains, after cultivation, the cells were washed in distilled water to make a standard sample of activated food-borne pathogenic bacteria strains, and the samples were injected after completion.

[0038] (2) Optimization of preparation conditions for enhanced reagents

[0039] In the present invention, gold nano-sol, silver nano-sol or other similar reagents can be selected as the reinforcing reagent. Wherein, gold nano-sol and silver nano-sol can be prepared by methods known to those skilled in the art. For example, gold nano-sol can be prepared by reducing chloroauric acid or its salts with sodium citrate, while silver nano-sol can be prepared by reducing soluble silver salt with NaBH4.

[0040] (3) Setting of Raman Spectrometer Injection Conditions

[0041] (4) Collection of Raman spectra of tested bacteria

[0042] The activated bacterial strain standard sample prepared i...

Embodiment 1

[0051]Example 1: Identification of four foodborne pathogens using surface-enhanced Raman spectroscopy: Staphylococcus aureus, Salmonella spp, Listeria monocytogenes and Escherichia coli (Escherichia coli), the process is:

[0052] Take the frozen strains of the above four pathogenic bacteria and inoculate them into TSB centrifuge tubes for activation, and activate them at 30-37°C for at least 24 hours. Scattered colonies were formed after 48 hours. Use a sterile inoculation loop to pick a single colony into a centrifuge tube filled with 4 mL of distilled water, vibrate and turbid, centrifuge in a refrigerated centrifuge at 8500rmp for 5 minutes, and repeat more than 3 times. Take the precipitate and wash it with water before injection. Take 500 μL of the gold nanosol prepared according to the above-mentioned optimization conditions and 300 μL of the above-mentioned bacteria standard sample and mix them in the injection bottle for 8 seconds, and measure 200mw by Raman, and get ...

Embodiment 2

[0053] Example 2: Using Surface Enhanced Raman Spectroscopy to Identify Unknown Foodborne Pathogenic Bacteria Samples

[0054] In this embodiment, according to the above method, 18 unknown food-borne pathogenic bacteria samples were sampled separately, and the obtained Raman spectrum was collected, and the spectral data were processed by SPSS software to obtain the cluster analysis results, and then the spectral data and cluster analysis The results were compared with the spectrograms and cluster analysis results of the aforementioned standard samples, and thus determined samples numbered 14, 16, samples 3, 4, 9, 10, 12, 13, 17, 18, 1, The samples of 5, 7, and 11, and the samples of 2, 6, 8, and 15 were Listeria monocytogenes, Salmonella, Escherichia coli, and Staphylococcus aureus, respectively, thus realizing the unknown food-borne pathogenic bacteria sample species Genus identification.

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Abstract

The invention relates to a method for identifying foodborne pathogen by surface enhanced Raman spectroscopy, which comprises the following steps: detecting foodborne pathogen by surface enhanced Raman spectroscopy by using gold or silver nano sol as an enhancing reagent, and carrying out cluster analysis on the detection result, thereby identifying the foodborne pathogen. The method provided by the invention has the advantages of high sensitivity, favorable selectivity, high detection speed and low cost, is easy to operate, can realize large-scale on-line detection of foodborne pathogen, and is widely applicable to the technical fields of food safety, environmental monitoring and the like.

Description

technical field [0001] The invention relates to a method for detecting food-borne pathogenic bacteria, in particular to a method for identifying food-borne pathogenic bacteria by using surface-enhanced Raman spectroscopy and cluster analysis. Background technique [0002] Food-borne pathogens are one of the main causes of food poisoning, which not only lead to the occurrence of various human diseases, but also seriously threaten people's health and cause economic losses. The World Health Organization estimates that billions of people around the world are infected with foodborne diseases every year. After the "9.11" incident in the United States, great attention has been paid to bioterrorism incidents around the world, so it is imperative to establish identification and detection methods for foodborne pathogens. Generally speaking, pathogenic bacteria causing foodborne diseases emerge in endlessly, but Salmonella, Escherichia coli, Staphylococcus aureus and Listeria monocyto...

Claims

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Application Information

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IPC IPC(8): C12Q1/04
CPCY02A50/30
Inventor 姚卫蓉王毅谦黄玉坤邵景东汪朋汪仕韬
Owner JIANGNAN UNIV
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