Non-diagnostic purpose rapid detection method for Escherichia coli O157:H7

A technology of Escherichia coli and Escherichia coli, which is applied in the field of biological detection, can solve problems that have not been reported, and achieve the effects of improving sensitivity, improving modification efficiency, and being simple, convenient and easy to prepare

Inactive Publication Date: 2018-07-20
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, gold-platinum-silica nanospheres (APS NPs) are simultaneously labeled with antibodies and enzymes to prepare signal transduction probe Abs. 2 / APS NPs / Invertase, applied to the detection of food-borne pathogens and other analytes have not been reported

Method used

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  • Non-diagnostic purpose rapid detection method for Escherichia coli O157:H7
  • Non-diagnostic purpose rapid detection method for Escherichia coli O157:H7
  • Non-diagnostic purpose rapid detection method for Escherichia coli O157:H7

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] (1) Fe 3 o 4 / SiO 2 Synthesis of Magnetic Nanoparticles with Core-Shell Structure

[0047] Synthesis of Fe by Solvothermal Method 3 o 4 Nanoparticles. The specific synthesis process is as follows: 0.5 g of poly(4-styrenesulfonic acid-co-maleic acid) sodium salt PSSMA was dissolved in 20 mL of ethylene glycol, and magnetically stirred to obtain a uniform orange solution. Add 0.54 g of ferric chloride hexahydrate and 1.5 g of anhydrous sodium acetate. Next, pour the obtained red-brown homogeneous liquid into a 100 mL hydrothermal reaction kettle, and react at 200° C. for 10 h. After cooling to room temperature, the obtained black precipitate was washed with ultrapure water at least three times under the action of an external magnetic field, and the volume was adjusted to 6 mL of ultrapure water for use.

[0048] Take 1 mL of the Fe prepared above 3 o 4 The solution was placed in an Erlenmeyer flask, 60mL of ethanol, 10mL of deionized water and 9mL of ammonia wate...

Embodiment 2

[0064] Example 2: Establishment of a method for rapid detection of Escherichia coli O157:H7 by signal transduction probe combined with immunomagnetic probe immuno-sandwich method

[0065] (1) Optimization of the amount of immunomagnetic probes used

[0066] The amount of immunomagnetic probes was changed to 30 μL, 35 μL, 40 μL, 50 μL and 55 μL respectively, and other experimental conditions were the same as in Example 1. Such as Figure 7 As shown, the concentration of glucose produced by sucrase hydrolysis increases continuously with the increase of the amount of capture probe, and the signal increases continuously, until the added amount reaches 50 μL, the concentration of glucose reaches the maximum, and the signal of PGM is also the strongest. As the amount continues to increase, the glucose concentration decreases instead. Therefore, the amount of capture probe added was chosen to be 50 μL.

[0067] (2) Sucrase and E.Coli O157:H7 labeled antibody modified on Au-Pt-SiO ...

Embodiment 3

[0071] Embodiment 3: Sensitivity detection

[0072] Under the detection condition of embodiment 1, have studied this detection method to 3.5 * 10 1 to 3.5×10 8 The detection results of E.Coli O157:H7 in the range of CFU / mL. The relationship between the PGM signal value and the concentration of E.Coli O157:H7 is as follows Figure 10 As shown, when the concentration of E.Coli O157:H7 is 3.5×10 2 CFU / mL to 3.5×10 6 When the CFU / mL changes, the difference between the glucose concentration and the logarithmic value of the concentration of E.Coli O157:H7 presents a good linear relationship. The linear equation is y=5.0724x-6.6421, the coefficient of variation R 2 =0.9863, and the detection limit of the method to the target bacteria E.Coli O157:H7 is 1.83×10 2 CFU / mL, the detection limit is higher than the traditional ELISA method (1.0×10 5 CFU / mL) is much lower. The concentration of E.Coli O157:H7 in the sample can be quantitatively obtained by linear regression equation...

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Abstract

The invention discloses a non-diagnostic purpose rapid detection method for Escherichia coli O157:H7. The rapid detection method comprises the following steps: combining an antibody with an enzyme byusing a Au-Pt-SiO2 composite nanosphere as a carrier to form a signal transduction probe; enriching and separating target substances in a sample by using the superparamagnetism of an immunomagnetic probe; and converting the signal of the detected target bacterium Escherichia coli O157:H7 into a glucose molecule signal, and reading the glucose molecule signal by a portable glucometer. The economic,simple, rapid and sensitive novel method for detecting Escherichia coli O157:H7 in foods is constructed on the basis of the glucometer and two nano-compounds capture probe and signal transduction probe.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a rapid detection method for non-diagnostic Escherichia coli O157:H7. Background technique [0002] Escherichia coli O157:H7 (Escherichia coli O157:H7, E.Coli O157:H7), is an important food-borne pathogenic bacteria, one of the top five major food-borne pathogenic bacteria. E.coli O157:H7 is mainly transmitted by contaminating beef, milk, chicken and its products, vegetables and other foods. When people are infected with E.Coli O157:H7, they will develop gastrointestinal diseases, and even develop thrombocytopenic purpura (TTP) in severe cases [8-10] . At present, the detection methods of E.Coli O157:H7 mainly include the national standard method (enrichment culture method) and various PCR methods, each with its own advantages and disadvantages. Therefore, it is of great practical significance to establish a better qualitative and quantitative new rapid detection ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N33/577
CPCG01N33/54346G01N33/56916G01N33/577
Inventor 赵广英窦文超叶玲娴
Owner ZHEJIANG GONGSHANG UNIVERSITY
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