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Enhanced Raman substrate for detecting streptomycete hyphae, and preparation and use methods thereof

A Raman substrate, Streptomyces technology, applied in Raman scattering, measuring devices, instruments, etc., can solve problems such as being unsuitable for the detection of large-scale samples, complex microbial serotypes, and cumbersome detection processes, and to promote miniaturization and Integration, shortened detection time, enhanced Raman spectrum

Active Publication Date: 2017-08-18
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection methods of microorganisms mainly include culture and dyeing observation method, immunological method, molecular biology method and mass spectrometry. Traditional microbial culture methods generally need to go through steps such as enrichment, separation and culture, staining or biochemical identification, and the whole detection process is cumbersome. , time-consuming, and heavy workload, it generally takes 4 to 7 days to get the test results. Immunological methods have the advantages of good specificity and high sensitivity, but due to the complexity of microbial serotypes, false positive results often occur in the test, and positive results need to be tested. Further tests confirmed
In recent years, molecular biology technology based on polymerase chain reaction (PCR) technology has developed very rapidly due to the advantages of strong specificity and high sensitivity, but some microorganisms will produce humic acid, which inhibits the PCR reaction, resulting in false positive results
The method of mass spectrometry has strong specificity and high sensitivity for microbial detection, but the cost is high, and it is not suitable for the detection of large batches of samples

Method used

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  • Enhanced Raman substrate for detecting streptomycete hyphae, and preparation and use methods thereof
  • Enhanced Raman substrate for detecting streptomycete hyphae, and preparation and use methods thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Preparation of nylon microporous filter membrane multilayer silicon dioxide silver-plated Raman detection substrate

[0037] 1. First, inject 1-100mL of 100-500nm diameter silica nanoparticle solution into a detachable filter with a diameter of 1-10cm through a syringe. The diameter of the detachable filter is 1-10cm and the filter hole diameter Assemble a multi-layer ordered structure on a nylon microporous filter membrane between 100-450 nm (smaller than the particle size of silica nanoparticles) to obtain a monodisperse silica nanoparticle layer;

[0038] 2. Inject 1-50 mL of the previously prepared gold nanoparticle solution with a particle size of 1-50nm into the filter, and filter it to the surface of the silica nanoparticle to obtain a gold nanoparticle layer;

[0039] 3. Take out the nylon microporous filter membrane covered with the gold nanoparticle layer, place it on a hot table and dry it, then immerse it in electroless silver electroplating solution A (...

Embodiment 2

[0040] Example 2: Detection of the Raman spectrum of the dye 4-ATP

[0041] 1. Cut the pre-prepared nylon microporous filter membrane multilayer silica silver-plated substrate into a size of 3mm×3mm;

[0042] 2. Use a micropipette to take 5μL, 10 -3 ~10 -12 The 4-ATP dye of M was dropped onto the above-mentioned substrate, and then the Raman spectroscopy of 4-ATP was detected by the confocal Raman microscope to obtain the Raman spectrum of the dye 4-ATP;

[0043] 3. After the reaction is completed, discard the used substrate. The substrate is a disposable substrate.

Embodiment 3

[0044] Example 3: Detection of the Raman spectrum of the dye R6G

[0045] 1. Cut the pre-prepared nylon microporous filter membrane multilayer silica silver-plated substrate into a size of 3mm×3mm;

[0046] 2. Use a micropipette to take 5μL, 10 -3 ~10 -12 The R6G dye of M is dropped onto the above-mentioned substrate, and then the Raman spectroscopy of R6G is detected by the confocal Raman microscope to obtain the Raman spectrum of the dye R6G;

[0047] 3. After the reaction is completed, discard the used substrate. The substrate is a disposable substrate.

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Abstract

The invention provides an enhanced Raman substrate for detecting streptomycete hyphae, preparation and use methods thereof. The detection substrate is composed of a filter membrane layer (1), a monodisperse nanometer particle layer (2) and a metallic coating (3) from bottom to top. The detection substrate is high in detection speed and sensitivity. The preparation process is as follows: 1) self-assembling on the filter membrane layer (1), thereby acquiring the monodisperse nanometer particle layer (2); 2) preparing a gold nanoparticle layer on the monodisperse nanometer particle layer (2); and 3) taking gold nanoparticles in the gold nanoparticle layer as a nucleus for growing the metallic coating (3), thereby acquiring the detection substrate. The preparation method is simple and fast. The detection substrate can be used for detecting if a detected sample contains the streptomycete hyphae; and the Raman spectrum of the sample solution is collected for detecting if the streptomycete hyphae are contained in the detected sample and detecting the content of the streptomycete hyphae. The method has wide application prospects in the fields of clinical detection, inspection quarantine, environment monitoring, and the like.

Description

Technical field [0001] The invention relates to an enhanced Raman substrate for detecting Streptomyces hyphae and a preparation and use method thereof, belonging to the field of bacteria detection. Background technique [0002] Nosiheptide, also known as Nosihitide, Nopeptidin or Nopeptomycin, is a sulfur-containing peptide antibiotic. It was first discovered by French scientists in the fermentation broth of Streptomyces actllosl ZS 40037 in 1961, and then obtained in the soil of Argentina. A strain that synthesizes Nosiheptide, it is sensitive to most Gram-positive bacteria, especially Staphylococcus aureus, Streptococcus and Clostridium weichii. By hindering the protein synthesis of bacteria, it inhibits the growth of bacteria, so it can Prevent and treat respiratory diseases and necrotizing enteritis. At present, many countries and regions (such as the European Union, Japan, Taiwan, China, etc.) allow nosiheptide to be used as a feed additive. In 1998, my country approved nos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 赵祥伟王德龙顾忠泽
Owner SOUTHEAST UNIV
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