CN108226135B - Composite membrane sensor and method for detecting vibrio parahaemolyticus by using same - Google Patents
Composite membrane sensor and method for detecting vibrio parahaemolyticus by using same Download PDFInfo
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Abstract
The invention provides a method for detecting vibrio parahaemolyticus by using an AU-PDMS composite membrane based on aptamer recognition. The sandwich structure detection sensor is established by fixing the specific recognition vibrio parahaemolyticus aptamer on the surface of the nanogold and modifying a Raman signal molecule as a signal probe, and fixing the specific recognition vibrio parahaemolyticus on an APT-AU-PDMS composite membrane formed by charge action as a detection substrate. The method has the advantages that the minimum detection limit of the vibrio parahaemolyticus can reach 12cfu/mL, the method can be applied to detection of the vibrio parahaemolyticus in the shrimp meat sample, and the result is accurate and reliable.
Description
Technical Field
The invention relates to the field of food safety detection, in particular to a method for detecting vibrio parahaemolyticus based on an aptamer-nanogold-PDMS composite membrane sensor.
Background
Vibrio parahaemolyticus, a halophilic bacterium widely present in marine products such as shrimps, crabs and shellfish, as well as in seawater and seabed sediments, has caused food poisoning events that surpass those caused by Salmonella. When people eat food infected by vibrio parahaemolyticus by mistake, acute gastroenteritis, diarrhea, septicemia and the like can be caused, and great harm is caused to the health of the people, and the outbreak of food-borne diseases caused by the vibrio parahaemolyticus is one of the important public health problems at present. Vibrio parahaemolyticus is widely distributed in seawater, seawater sediments, fish, shrimp, crab, shellfish and other aquatic products near the coast, and a large number of aquatic products are reported to be polluted by the Vibrio parahaemolyticus in coastal cities and inland areas such as Guangdong, Fujian, Shandong, Zhejiang, Jiangsu, Shanghai and the like in China, and food poisoning reports caused by the Vibrio parahaemolyticus are also rare. In view of the above, China sets the limit standard for the quantity of vibrio parahaemolyticus in aquatic products and aquatic seasoning products, wherein n is 5, c is 1, M is 100cfu/g (mL), and M is 1000cfu/g (mL). Therefore, the establishment of a sensitive, rapid and economic vibrio parahemolyticus detection technology has important significance for guaranteeing food safety.
At present, the conventional detection methods for vibrio parahaemolyticus mainly comprise the traditional microbial detection technology, molecular biology technology, immunological detection method and the like. The traditional detection method is long in time consumption, complex in operation and expensive in cost, and cannot meet the requirements of rapidness, simplicity and economy for food safety detection; although the molecular biology technology can shorten the detection time, the total DNA of bacteria needs to be extracted in the early stage, the detection object is the target DNA rather than the bacteria, the detection result needs to be subjected to data conversion, and certain equipment and cost are needed; the immunological method has the advantages of strong specificity, high sensitivity, easy observation and the like, but the antibody as a recognition molecule is easily influenced by external conditions, and has low stability and high price. In order to overcome the defects, the development of a sensitive, specific, rapid and economic detection means for monitoring the vibrio parahaemolyticus is urgently needed to ensure the food safety of people.
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 adopts a sandwich method to carry out Raman spectroscopy to detect pathogenic bacteria, but the system is a solution, the detection is limited by a plurality of conditions, and the steps of centrifugation and the like are required, so the operation is not favorable for rapid detection.
CN106093005A provides a latent fingerprint high-definition recognition method based on lysozyme Raman spectrum imaging, the method collects fingerprints on a substrate and then adopts a sandwich method to carry out Raman spectrum imaging, but the substrate requirement is too high, incubation is required under the conditions of specified temperature and humidity, and therefore, the method has certain limitation.
Disclosure of Invention
In view of the above problems in the prior art, the present application provides a composite membrane sensor and a method for detecting vibrio parahaemolyticus thereof. The method has the advantages of high sensitivity, strong specificity, convenient operation, economy and practicality, and has wide application prospect in the field of food safety detection.
The technical scheme of the invention is as follows:
a composite membrane sensor is characterized in that an aptamer-nanogold-composite membrane sensor is constructed by fixing a specificity recognition vibrio parahaemolyticus aptamer on the surface of nanogold and modifying a Raman signal molecule to serve as a signal probe, and fixing the specificity recognition vibrio parahaemolyticus on an APT-AU-PDMS composite membrane formed by charge action to serve as a detection substrate.
The specific preparation method of the composite membrane sensor comprises the following steps:
(1) immersing the PDMS membrane in piranha solution for 30-90s at normal temperature, performing high-temperature treatment with APTES solution at 70 ℃, repeatedly performing ultrasonic treatment with deionized water for three times, and immersing in nano gold solution to obtain AU-PDMS membrane;
(2) under the conditions of a shaking table at 37 ℃ and 150-200 rpm, immersing the AU-PDMS membrane in 40nM TE buffer solution of an aptamer capable of specifically recognizing the vibrio parahaemolyticus, and incubating for 12-48h to obtain an APT-AU-PDMS composite membrane;
(3) adding equal volume of 1mM TCEP solution into 10 μ L of 10 μ M aptamer solution, standing for half an hour, incubating 1mL of nanogold solution with the aptamer solution for specifically recognizing Vibrio parahaemolyticus for 12h, adding 10-30 μ L of 10% SDS solution, aging at 37 ℃ for 8-18h at the rotation speed of 140-;
(4) marking a Raman signal molecule 4-MBA on the surface of the nano gold compounded with the aptamer for specifically recognizing the vibrio parahaemolyticus to serve as a signal probe;
(5) an APT-AU-PDMS composite membrane is used as a detection substrate, and a PDMS-nanogold-5' end sulfhydrylation vibrio parahaemolyticus aptamer sensor is established.
The Raman signal molecule 4MBA is 1590cm-1Characteristic peak of (c).
Wherein, PDMS is named as polydimethylsiloxane, the thickness of PDMS membrane is about 1mm, APT is named as aptamer, TCEP solution is named as tris (2-carboxyethyl) phosphine solution.
The beneficial technical effects of the invention are as follows:
the detected object is the vibrio parahaemolyticus thallus, the adopted recognition molecule is an aptamer capable of specifically binding to the whole vibrio parahaemolyticus, and recognition and capture are realized through the cooperation of the spatial structure of the aptamer and the vibrio parahaemolyticus. When the vibrio parahaemolyticus exists in the system, the signal probe is specifically combined with the vibrio parahaemolyticus and is fixed on the detection substrate, and a Raman signal is generated.
Measurement at 633nm as the emission light source to 1590cm-1And detecting the intensity of the Raman signal by monitoring the change of the Raman signal. By detecting the vibrio parahaemolyticus with different concentrations, a standard curve is established, and the aim of quantitatively detecting the vibrio parahaemolyticus-containing sample is fulfilled. The specific detection principle is shown in fig. 1.
The principle of the present application is different from CN105352933A and CN106093005A in that a solid nano-aptamer composite substrate is prepared, and simultaneously, the sensitivity is higher. Based on such difference, compared with the two technologies, the application firstly fixes the aptamer on the PDMS film, so that a system with low cost, uniformity, stability and convenient operation is formed, and meanwhile, the sensitivity is better.
Compared with an immunoassay method in which an antibody is used as an identification element, the method takes the aptamer as the identification element, has the advantages of good aptamer stability, low preparation cost, easy labeling, no influence on the activity of the aptamer after labeling, high affinity and high selectivity on target thalli, and greatly improves the detection accuracy.
The invention takes the whole bacteria as the detection object, and compared with the detection of bacterial DNA, the method is more accurate, convenient and fast. The invention combines the aptamer and the AU-PDMS composite membrane to construct the aptamer-nanogold-PDMS composite membrane sensor which is used for detecting the vibrio parahaemolyticus, can greatly shorten the detection time, has the advantages of quickness, economy and the like, and has wide application prospect.
Compared with a solid substrate, the AU-PDMS detection substrate has the advantages of economy and good light stability, can realize high-sensitivity detection on the vibrio parahaemolyticus, and has the detection limit of 12 cfu/mL.
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FIG. 1 is a schematic view of the method for detecting Vibrio parahaemolyticus by the composite membrane sensor of the present invention;
FIG. 2 is an SEM image of an AU-PDMS composite membrane;
FIG. 3 is a spectrum of Raman intensity changes caused by different concentrations of Vibrio parahaemolyticus;
FIG. 4 is a graph showing the linear relationship between the relative Raman intensity and the concentration of Vibrio parahaemolyticus.
Detailed Description
The present invention will be described in detail with reference to the drawings and examples, but the present invention is not limited thereto.
Example 1: preparation of composite membrane sensor
(1) Preparing a nano gold solution:
0.5ml of 1% chloroauric acid aqueous solution is taken, 49ml of deionized water is added, boiling is carried out for 5-10 minutes (120 rotating speeds), then 0.5ml of 5% sodium citrate is rapidly added, and the nano-gold solution is obtained after the solution reacts to generate wine red. And (3) centrifuging 1ml of the prepared nanogold solution at 12000RPM for 20min, removing 750 mu L of supernatant, then resuspending to obtain a concentrated nanogold solution, and storing the concentrated nanogold solution in a refrigerator at 4 ℃.
(2) Preparing an APT-AU-PDMS composite membrane:
host in PDMS: the adjuvant (i.e., polydimethylsiloxane prepolymer: curing agent) was 10: 1. The main agent and the fixing agent are put into a test tube and fully stirred. Stirring until the size of the bubbles in the colloid is uniform and no obvious large bubbles exist, which indicates that the main agent and the fixing agent of the PDMS are uniformly mixed. Slowly pouring 0.285ml of mixed colloid into a plastic culture dish, flatly spreading the colloid to a substrate, putting the culture dish into a vacuum drying oven, and vacuumizing for 1h at normal temperature until all bubbles in the colloid disappear. And then putting the culture dish into an oven, drying at the temperature of 70 ℃ overnight, and obtaining a pure PDMS membrane after the PDMS colloid is completely solidified.
Cutting the prepared PDMS membrane into 0.5 x 0.5cm pieces by a blade, placing the PDMS pieces into a 1.5ml centrifuge tube, treating the PDMS pieces with piranha solution at 40 ℃ for 60s, repeatedly washing the PDMS pieces with deionized water for three times, treating the hydroxylated PDMS membrane with 5% APTES ethanol solution at 70 ℃ for 3h, repeatedly washing the prepared aminated PDMS membrane with deionized water for three times by ultrasonic, and drying the membrane with nitrogen. Incubating a proper amount of nano gold solution and the prepared aminated PDMS membrane overnight, repeatedly washing with deionized water to obtain an AU-PDMS membrane, immersing the AU-PDMS membrane in a proper amount of aptamer solution, and reacting at 37 ℃ for 24H at the rotating speed of 180 ℃ of a shaking table to obtain the APT-AU-PDMS composite membrane. As shown in particular in fig. 2.
(3) Preparation of a Raman signal probe:
mixing and standing equivalent vibrio parahaemolyticus aptamer (10 mu M) and TECP (1mM) for half an hour, adding 40 mu L of vibrio parahaemolyticus aptamer (10 mu M) into 1ml of nano gold solution, and placing the solution into a shaking table at the rotating speed of 150 for reaction for 12 hours. Then 20 mul 10% SDS solution is added into the solution, and the solution is aged for 12h at 37 ℃ under the rotation speed of 150, 0.5mol/L NaCl solution is added successively in the aging process to make the final concentration of NaCl reach 0.15mol/L, and after each addition of NaCl solution, the solution is treated by ultrasonic treatment for about 10 s. After aging, centrifuging at 12000r/min at 25 deg.C for 30min, discarding supernatant, dissolving precipitate with ultrapure water, repeating twice to remove aptamer not connected to nanogold, and storing at 4 deg.C for use. 1mL of the prepared solution was added with 0.1mL of 4-MBA (1mM) ethanol solution, and the mixture was placed in a shaker at 150rpm for 12 hours. So as to obtain the Raman signal probe molecule.
(4) Detection of Vibrio parahaemolyticus:
taking 500 mu L of vibrio parahaemolyticus liquid with different concentrations, placing the vibrio parahaemolyticus liquid and the APT-AU-PDMS composite membrane in a shaker at the rotating speed of 150 ℃ for incubation for 1h, washing twice by PBS buffer solution to remove the liquid physically adsorbed on the APT-AU-PDMS composite membrane, taking 500 mu L of signal probe solution and the washed APT-AU-PDMS composite membrane, placing the APT-AU-PDMS composite membrane in a shaker at the rotating speed of 150 ℃ for incubation for 1h, taking out the APT-AU-PDMS composite membrane, and washing twice by PBS buffer solution to remove the redundant signal probe physically adsorbed on the APT-AU-PDMS composite membrane. When vibrio parahaemolyticus exists in the system, the specificity of the signal probe is combined with the vibrio parahaemolyticus, and the raman intensity of the signal molecule is enhanced, so that the detection of the vibrio parahaemolyticus is realized by monitoring the change of the raman intensity, and the specific structure is shown in fig. 3.
Example 2: establishment of standard detection curve of vibrio parahaemolyticus
Adding 500 μ L of vibrio parahaemolyticus liquid with different concentrations into APT-AU-PDMS composite membrane, incubating for 1h at 37 deg.C with 150rpm shaking table, and incubatingAnd then washing twice with PBS buffer solution, taking 500 mu L of signal probe solution and placing the signal probe solution and the signal probe solution in a shaking table at the rotating speed of 150 ℃ for incubation for 1h, then washing twice with the APT-AU-PDMS composite membrane by using the PBS buffer solution, and measuring the Raman intensity (633 nm of excitation light source). As shown in FIG. 4, Vibrio parahaemolyticus is at 1X 101-1×105The concentration of cfu/mL is in the range of 1590cm-1The relative Raman intensity is in a good linear relation, the linear equation is that y is 588.0x-495.8(R2 is 0.990), and the lowest detection limit is 12 cfu/mL. As shown in particular in fig. 4.
Test example 1: detection of vibrio parahaemolyticus in shrimp meat sample
Fresh shrimp was purchased from a local supermarket, the shell was opened by aseptic manipulation, a sample of shrimp meat was minced at 25g and homogenized with 225mL of 3% sodium chloride alkali peptone water for 10 min. Because the substrate has stronger stability, 500 mu L of mixed and homogenized mixture can be directly taken out to a centrifuge tube as an actual sample, 4 samples are taken in total, vibrio parahaemolyticus with known concentration (the concentration is obtained by a flat plate counting method) is added, then the mixture and the APT-AU-PDMS composite membrane are placed on a 37 ℃ shaking table at 150rpm for incubation for 1h, then PBS buffer solution is used for cleaning twice, 500 mu L of signal probe solution is taken and placed on a 37 ℃ shaking table at 150rpm for incubation for 1h, then the PBS buffer solution is used for cleaning the APT-AU-PDMS composite membrane twice, and the Raman intensity (with 633nm excitation light source) is measured. The results are shown in table 1, the detection results obtained by the method of the invention have no significant difference from the plate counting method, and the recovery rate is between 94% and 105.3%.
TABLE 1 detection results of Vibrio parahaemolyticus in shrimp meat samples
Detection example 2: sensitivity of the detection method of the invention
A sample of No.1 from test example 1 (i.e., with the addition of 3.0X 10)2cfu/mL of a test solution of Vibrio parahaemolyticus) was diluted 5 times, 10 times, and 20 times, respectively, and the Raman intensity (633 nm excitation light source) was measured using the composite sensor of the present invention. The results are shown in Table 2.
TABLE 2
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (2)
1. The application of the composite membrane sensor in detecting the vibrio parahaemolyticus is characterized in that: the composite membrane sensor is constructed by fixing an aptamer for specifically recognizing vibrio parahaemolyticus on the surface of nanogold and modifying a Raman signal molecule to serve as a signal probe, and fixing the aptamer for specifically recognizing vibrio parahaemolyticus on an APT-AU-PDMS composite membrane formed by charge action to serve as a detection substrate; the detection comprises the steps of taking an APT-AU-PDMS composite membrane, adding vibrio parahaemolyticus liquid, placing the APT-AU-PDMS composite membrane in a shaker at the rotating speed of 150rpm at 37 ℃ for incubation for 1h, then cleaning the APT-AU-PDMS composite membrane by using PBS buffer solution, taking 500 mu L of signal probe solution and placing the signal probe solution in the shaker at the rotating speed of 150rpm at 37 ℃ for incubation for 1h, then cleaning the APT-AU-PDMS composite membrane by using the PBS buffer solution;
the specific preparation method of the aptamer-nanogold-composite membrane sensor comprises the following steps:
(1) immersing the PDMS membrane in piranha solution for 30-90s at normal temperature, performing high-temperature treatment with APTES solution at 70 ℃, repeatedly performing ultrasonic treatment with deionized water for three times, and immersing in nanogold solution to obtain AU-PDMS membrane;
(2) under the conditions of a shaking table at 37 ℃ and 150-200 rpm, immersing the AU-PDMS membrane in 40nM TE buffer solution of an aptamer capable of specifically recognizing the vibrio parahaemolyticus, and incubating for 12-48h to obtain an APT-AU-PDMS composite membrane;
(3) adding equal volume of 1mM TCEP solution into 10 μ L of 10 μ M aptamer solution, standing for half an hour, incubating 1mL of nanogold solution with the aptamer solution for specifically recognizing Vibrio parahaemolyticus for 12h, adding 10-30 μ L of 10% SDS solution, aging at 37 ℃ for 8-18h at the rotation speed of 140-;
(4) marking a Raman signal molecule 4-MBA on the surface of the nano gold compounded with the aptamer for specifically recognizing the vibrio parahaemolyticus to serve as a signal probe;
(5) an APT-AU-PDMS composite membrane is used as a detection substrate, and a PDMS-nanogold-5' end sulfhydrylation vibrio parahaemolyticus aptamer sensor is established.
2. Use according to claim 1, characterized in that: the Raman signal molecule is 4MBA and is 1590cm-1There is a characteristic peak.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104730062A (en) * | 2015-04-16 | 2015-06-24 | 苏州大学 | Transparent flexible surface-enhanced Roman spectroscopy substrate (SERS) and preparation method and application thereof |
| CN105352933A (en) * | 2015-09-29 | 2016-02-24 | 江南大学 | Method for detection of vibrio parahaemolyticus in food on basis of aptamer identification surface enhanced Raman spectrum |
| CN105755463A (en) * | 2016-03-07 | 2016-07-13 | 东南大学 | Method for preparing hydrophobic surface solid monolayer uniform SERS substrate |
| CN106645087A (en) * | 2016-12-22 | 2017-05-10 | 东南大学 | PDMS-base single-layer SERS substrate not requiring surface modification and preparation method thereof |
| CN107255631A (en) * | 2017-05-25 | 2017-10-17 | 西南交通大学 | A kind of Raman spectrum base based on PDMS sponges and preparation method thereof |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104730062A (en) * | 2015-04-16 | 2015-06-24 | 苏州大学 | Transparent flexible surface-enhanced Roman spectroscopy substrate (SERS) and preparation method and application thereof |
| CN105352933A (en) * | 2015-09-29 | 2016-02-24 | 江南大学 | Method for detection of vibrio parahaemolyticus in food on basis of aptamer identification surface enhanced Raman spectrum |
| CN105755463A (en) * | 2016-03-07 | 2016-07-13 | 东南大学 | Method for preparing hydrophobic surface solid monolayer uniform SERS substrate |
| CN106645087A (en) * | 2016-12-22 | 2017-05-10 | 东南大学 | PDMS-base single-layer SERS substrate not requiring surface modification and preparation method thereof |
| CN107255631A (en) * | 2017-05-25 | 2017-10-17 | 西南交通大学 | A kind of Raman spectrum base based on PDMS sponges and preparation method thereof |
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