Composite membrane sensor and method thereof for detecting vibrio parahemolyticus

A hemolytic vibrio and composite membrane technology, applied in the direction of Raman scattering, material excitation analysis, etc., can solve the problems of high substrate requirements, unfavorable rapid detection, cumbersome operation, etc., achieve low preparation cost, shorten detection time, The effect of broad application prospects

Active Publication Date: 2018-06-29
JIANGNAN UNIV
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  • Description
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  • Application Information

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Problems solved by technology

[0004] CN105352933A provides a method for detecting Vibrio parahaemolyticus in food based on aptamer recognition surface-enhanced Raman spectroscopy. The method prepares a silver-coated gold core-shell substrate and uses a sandwich method for Raman spectroscopy to detect pathogenic bacteria , but because the system is a solution, the detection is limited by many conditions, and centrifugation and other steps are required, which is not conducive to rapid detection.
[0005] CN106093005A provides a latent fingerprint high-definition identification method based on lysozyme Raman spectrum imaging. The method collects fingerprints on the substrate and uses the sandwich method to perform Raman spectroscopy for fingerprint imaging. , Incubated under humidity conditions, so there are certain limitations

Method used

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  • Composite membrane sensor and method thereof for detecting vibrio parahemolyticus
  • Composite membrane sensor and method thereof for detecting vibrio parahemolyticus
  • Composite membrane sensor and method thereof for detecting vibrio parahemolyticus

Examples

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Effect test

Embodiment 1

[0029] Example 1: Preparation of composite membrane sensor

[0030] (1) Preparation of nano gold solution:

[0031] Take 0.5ml of 1% chloroauric acid aqueous solution, add 49ml of deionized water and boil for 5-10 minutes (120 rotation speed), then quickly add 0.5ml of 5% sodium citrate, and wait until the solution reacts to produce wine red to obtain a nano gold solution. Take 1ml of the prepared nano-gold solution, centrifuge at 12000RPM for 20 minutes, remove 750 μL of supernatant and resuspend to obtain a concentrated nano-gold solution, and store it in a refrigerator at 4°C.

[0032] (2) Preparation of APT-AU-PDMS composite membrane:

[0033] The main agent in PDMS: auxiliary agent (ie, polydimethylsiloxane prepolymer: curing agent) = 10:1. Put the main agent and fixative into the test tube and stir thoroughly. Stir until the size of the bubbles in the colloid is uniform, and there are no obvious large bubbles, it means that the main agent and fixative of PDMS have been mixed e...

Embodiment 2

[0039] Example 2: Establishment of a standard detection curve for Vibrio parahaemolyticus

[0040] Take the APT-AU-PDMS composite membrane, add 500μL of Vibrio parahaemolyticus solution of different concentrations, incubate for 1h on a shaker at 37℃ at 150 speed, then wash twice with PBS buffer, and then take 500μL of signal probe solution Instead of placing it on a 37°C shaker and incubating at 150 speed for 1 hour, the APT-AU-PDMS composite membrane was washed twice with PBS buffer, and the Raman intensity (excitation light source 633nm) was measured. Such as Figure 4 As shown, Vibrio parahaemolyticus at 1×10 1 -1×10 5 cfu / mL concentration range and 1590cm -1 The relative Raman intensity showed a good linear relationship, the linear equation was y=588.0x-495.8 (R2=0.90), and the lowest detection limit was 12cfu / mL. Specific as Figure 4 Shown.

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Abstract

The invention provides a method for detecting vibrio parahemolyticus by an AU-PDMS composite membrane on the basis of aptamer identification. A specific recognition vibrio parahemolyticus aptamer is fixed on the surface of nanogold, a Raman signal module is modified as a signal probe, the specific recognition vibrio parahemolyticus is fixed on an APT-AU-PDMS composite membrane formed by a charge effect to serve as a detection substrate, and a 'sandwich' structure detection sensor is established. The lowest detection limit of the method for the vibrio parahemolyticus can be 12cfu/mL, in addition, the method can be applied to the detection of the vibrio parahemolyticus in shrimp samples, and a result is accurate and reliable.

Description

Technical field [0001] The invention relates to the field of food safety detection, in particular to a method for detecting Vibrio parahaemolyticus based on an aptamer-nano-gold-PDMS composite film sensor. Background technique [0002] Vibrio parahaemolyticus, as a kind of halophilic bacteria widely present in seafood such as shrimps, crabs, shellfish, sea water and seabed sediments, has caused food poisoning incidents that have surpassed Salmonella poisoning incidents. When people accidentally eat food infected by Vibrio parahaemolyticus, it can cause acute gastroenteritis, diarrhea, sepsis, etc., and cause great harm to health. The outbreak of food-borne diseases caused by Vibrio parahaemolyticus is the current One of the important public health problems facing. Vibrio parahaemolyticus is widely distributed in coastal seawater, seawater sediments, fish, shrimp, crab, shellfish and other aquatic products, in my country’s coastal cities and inland such as Guangdong, Fujian, Shan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
CPCG01N21/65
Inventor 段诺沈默斐王周平吴世嘉邹颖
Owner JIANGNAN UNIV
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