Magnetic fluorescent probe-based food escherichia coli colony visual detection and automatic counting method

A technology of Escherichia coli and fluorescent probes, applied in the field of microbial detection, can solve the problems of low reproducibility, time-consuming, and large randomness of results in immunoassays and molecular biology methods

Active Publication Date: 2019-09-20
JIANGSU UNIV
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Problems solved by technology

The traditional biochemical assay method realizes the detection of E. coli according to the characteristics of microbial growth, which is time-consuming, easily affected by the growth environment and has low accuracy
Immunoassay and molecular biology methods have disadvantages such as low reproducibility and high cost
Moreover, the above methods cannot realize the identification and counting of Escherichia coli at the same time
[0004] Colony most probable number method and

Method used

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  • Magnetic fluorescent probe-based food escherichia coli colony visual detection and automatic counting method
  • Magnetic fluorescent probe-based food escherichia coli colony visual detection and automatic counting method
  • Magnetic fluorescent probe-based food escherichia coli colony visual detection and automatic counting method

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Embodiment 1

[0047] In the present invention, the bacterial strains in the milk are solid-state cultured at first to obtain the colony plate of the bacterial strain; then the prepared specific magnetic fluorescent probe is added to the colony plate, and the magnetic fluorescent probe that does not react with E. coli is magnetically separated; Plate image; the final image processing realizes the automatic counting of E. coli colonies.

[0048] Step 1, magnetic nanosphere Fe 3 o 4 @SiO 2 - Preparation of cDNA:

[0049] Add 1.08g of ferric chloride hexahydrate, 1.8g of sodium acetate trihydrate and 0.25g of trisodium citrate into 50mL of ethylene glycol in sequence, and stir magnetically for 9 hours at 40°C to form a uniform and transparent solution, then transfer it to the reaction kettle in 200°C for 10 h, and finally the obtained mixture was washed three times with deionized water and ethanol, magnetically separated to obtain a black product, and vacuum dried at 60°C for 10 h to obtain ...

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Abstract

The invention belongs to the technical field of microbiological detection and relates to a magnetic fluorescent probe-based food escherichia coli colony visual detection and automatic counting method. According to the method, magnetic fluorescent probes are prepared, bacterial strain colonies in a food are cultured; the magnetic fluorescent probes are added into food bacterial strain colony flat plates, and the food bacterial strain colony flat plates are put into a portable fluorescence imager, so that bacterial colony fluorescence flat plate images are obtained; when escherichia coli exists in the colonies, specific fluorescent carbon quantum dots are adsorbed on the escherichia coli colonies, the escherichia coli colonies display fluorescence, otherwise, no fluorescence is displayed, and therefore, the visual identification of the escherichia coli colonies is realized; the obtained escherichia coli colony fluorescence flat plate images are inputted to a computer program so as to be processed, so that the automatic counting of the escherichia coli is realized. The escherichia coli colonies are positioned through the specific fluorescent probes, so that the fluorescence visual detection of the escherichia coli colonies is realized; and the content of the escherichia coli can be accurately calculated in real time.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, and in particular relates to a method for visual detection and automatic counting of food Escherichia coli colonies based on a magnetic fluorescent probe. Background technique [0002] Food can be contaminated by Escherichia coli in different ways. Under the action of E. coli, the food will be spoiled and deteriorate, lose its proper nutrients, and affect its edibility and safety. After people eat food contaminated by E. coli, it may cause extraintestinal infection, acute diarrhea, dysentery, cholera or typhoid and even severe sepsis. It seriously endangers human health and safety, and is an internationally recognized indicator bacteria for health monitoring. [0003] Existing techniques for detecting Escherichia coli mainly include traditional biochemical assays, immunoassays, and molecular biology methods. Traditional biochemical assays are based on the characteristics of micr...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/58G06T7/136G06T7/187G06T5/00
CPCG01N33/56916G01N33/582G06T7/136G06T7/187G06T5/002G01N2333/245G06T2207/30242
Inventor 邹小波胡雪桃石吉勇李文亭李艳肖黄晓玮李志华徐艺伟李亚惠
Owner JIANGSU UNIV
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