Microfluidic chip, detector, detection method and application for detecting bacteria

Abstract

The invention provides a microfluidic chip for detecting bacteria, a bacteria detector including the microfluidic chip, a method and an application in the technical field of bacteria detection. The microfluidic chip includes a sampling port, a reaction chamber and a chip channel; wherein, the sampling port and the reaction chamber are connected through a chip channel; the reaction chamber is pre-coated with an initiator chain and an aptamer; The microfluidic chip includes at least two layers, and the chip materials on at least one side of the reaction chamber are all visible materials. The invention provides a microfluidic chip and a detector for detecting food-borne germs, which are small in size, low in cost, convenient for mass production and easy to carry, and are based on microfluidic chip technology and hybridization chain reaction (HCR) and G- tetramer-mediated H 2 O 2 The color development method is combined, the operation is simple, the detection sensitivity is greatly improved, and the detection result is visible to the naked eye, which is suitable for commercial promotion and application.

Description

technical field

[0001] The invention relates to the technical field of rapid detection of bacteria, in particular to a microfluidic chip, a detector, a detection method and application for rapid detection of bacteria. Background technique

[0002] As an important factor in causing food-borne diseases, food-borne pathogens have attracted extensive attention from all walks of life. The pathogenic bacteria that usually cause foodborne diseases mainly include the following: pathogenic Escherichia coli, Salmonella, Shigella, Listeria monocytogenes, Vibrio parahaemolyticus, Streptococcus hemolyticus , Bacillus cereus and Staphylococcus aureus. Pathogenic bacteria in food can cause food safety concerns. These bacteria invade the body and produce toxins that can make people sick. Therefore, rapid detection of foodborne pathogens is crucial for the effective control of related diseases.

[0003] At present, the detection methods of foodborne bacteria mainly include conventional d...

Examples

Embodiment 1

[0072] The above method and device are used to detect the content of pathogenic Escherichia coli.

[0073] Pathogenic Escherichia coli: BNCC 133264, purchased from Beina Biological Company.

[0074] The sequence of the priming chain is shown in SEQ ID No. 1, which was synthesized by Shanghai Sangon Biological Company.

[0075] The aptamer sequence is shown in SEQ ID No. 2, which was synthesized by Shanghai Sangon Biological Company.

[0076] Reagents are H1 and H2 chains for the hybridization chain reaction, and hemin. The sequence of the H1 chain is shown in SEQ ID No. 3, and the sequence of H2 is shown in SEQ ID No. 4, both of which were synthesized by Shanghai Sangong Company. Hemin has a CAS# of 16009-13-5 and was purchased from SigmaAldrich.

[0077] According to the detection steps described in this application, after reaching S3, it was observed that the color of the reagent in the reaction chamber reached the fifth-grade green, and the content of pathogenic E. 6 Th...

Embodiment 2

[0082] The content detection of Bacillus cereus is carried out by using the above method and device.

[0083] Bacillus cereus: BNCC 336744, purchased from Beina Bio.

[0084] The sequence of the priming chain is shown in SEQ ID No. 1, which was synthesized by Shanghai Sangon Biological Company.

[0085] The aptamer sequence is shown in SEQ ID No. 5, which was synthesized by Shanghai Sangon Biotechnology Co., Ltd.

[0086] Reagents are H1 and H2 chains for the hybridization chain reaction, and hemin. The sequence of the H1 chain is shown in SEQ ID No. 6, and the sequence of the H2 chain is shown in SEQ ID No. 7, both of which were synthesized by Shanghai Sangong Company. Hemin was the same as in Example 1.

[0087] According to the detection steps described in this application, after S3, it is observed that the color of the reagent in the reaction chamber reaches the second-level green, and the content of Bacillus cereus determined after comparing with the colorimetric card ...

Embodiment 3

[0092] The above method and device are used to detect the content of Staphylococcus aureus.

[0093] Staphylococcus aureus: BNCC 186335, purchased from Beina Bio.

[0094] The sequence of the priming chain is shown in SEQ ID No. 1, which was synthesized by Shanghai Sangon Biological Company.

[0095] The aptamer sequence is shown in SEQ ID No. 8, which was synthesized by Shanghai Sangon Biological Company.

[0096] Reagents are H1 and H2 chains for the hybridization chain reaction, and hemin. The sequence of the H1 chain is shown in SEQ ID No. 9, and the sequence of the H2 chain is shown in SEQ ID No. 10, both of which were synthesized by Shanghai Sangong Company. Hemin was the same as in Example 1.

[0097] According to the detection steps described in this application, after reaching S3, it is observed that the color of the reagent in the reaction chamber reaches the sixth-grade green, and the content of Staphylococcus aureus determined after comparing with the colorimetr...

technical field

[0001] The invention relates to the technical field of rapid detection of bacteria, in particular to a microfluidic chip, a detector, a detection method and application for rapid detection of bacteria. Background technique

[0002] As an important factor in causing food-borne diseases, food-borne pathogens have attracted extensive attention from all walks of life. The pathogenic bacteria that usually cause foodborne diseases mainly include the following: pathogenic Escherichia coli, Salmonella, Shigella, Listeria monocytogenes, Vibrio parahaemolyticus, Streptococcus hemolyticus , Bacillus cereus and Staphylococcus aureus. Pathogenic bacteria in food can cause food safety concerns. These bacteria invade the body and produce toxins that can make people sick. Therefore, rapid detection of foodborne pathogens is crucial for the effective control of related diseases.

[0003] At present, the detection methods of foodborne bacteria mainly include conventional d...

Examples

Embodiment 1

[0072] The above method and device are used to detect the content of pathogenic Escherichia coli.

[0073] Pathogenic Escherichia coli: BNCC 133264, purchased from Beina Biological Company.

[0074] The sequence of the priming chain is shown in SEQ ID No. 1, which was synthesized by Shanghai Sangon Biological Company.

[0075] The aptamer sequence is shown in SEQ ID No. 2, which was synthesized by Shanghai Sangon Biological Company.

[0076] Reagents are H1 and H2 chains for the hybridization chain reaction, and hemin. The sequence of the H1 chain is shown in SEQ ID No. 3, and the sequence of H2 is shown in SEQ ID No. 4, both of which were synthesized by Shanghai Sangong Company. Hemin has a CAS# of 16009-13-5 and was purchased from SigmaAldrich.

[0077] According to the detection steps described in this application, after reaching S3, it was observed that the color of the reagent in the reaction chamber reached the fifth-grade green, and the content of pathogenic E. 6 Th...

Embodiment 2

[0082] The content detection of Bacillus cereus is carried out by using the above method and device.

[0083] Bacillus cereus: BNCC 336744, purchased from Beina Bio.

[0084] The sequence of the priming chain is shown in SEQ ID No. 1, which was synthesized by Shanghai Sangon Biological Company.

[0085] The aptamer sequence is shown in SEQ ID No. 5, which was synthesized by Shanghai Sangon Biotechnology Co., Ltd.

[0086] Reagents are H1 and H2 chains for the hybridization chain reaction, and hemin. The sequence of the H1 chain is shown in SEQ ID No. 6, and the sequence of the H2 chain is shown in SEQ ID No. 7, both of which were synthesized by Shanghai Sangong Company. Hemin was the same as in Example 1.

[0087] According to the detection steps described in this application, after S3, it is observed that the color of the reagent in the reaction chamber reaches the second-level green, and the content of Bacillus cereus determined after comparing with the colorimetric card ...

Embodiment 3

[0092] The above method and device are used to detect the content of Staphylococcus aureus.

[0093] Staphylococcus aureus: BNCC 186335, purchased from Beina Bio.

[0094] The sequence of the priming chain is shown in SEQ ID No. 1, which was synthesized by Shanghai Sangon Biological Company.

[0095] The aptamer sequence is shown in SEQ ID No. 8, which was synthesized by Shanghai Sangon Biological Company.

[0096] Reagents are H1 and H2 chains for the hybridization chain reaction, and hemin. The sequence of the H1 chain is shown in SEQ ID No. 9, and the sequence of the H2 chain is shown in SEQ ID No. 10, both of which were synthesized by Shanghai Sangong Company. Hemin was the same as in Example 1.

[0097] According to the detection steps described in this application, after reaching S3, it is observed that the color of the reagent in the reaction chamber reaches the sixth-grade green, and the content of Staphylococcus aureus determined after comparing with the colorimetr...

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