Test strip for detecting wide spectrum type salmonella in food and preparation and use method thereof

Abstract

The invention, which relates to the field of biotechnology, particularly discloses a test strip for detecting wide spectrum type salmonella in food and a preparation and use method thereof. The test strip comprises a PVC bottom plate, a nitrocellulose film bonded to the middle section of at least one surface of the PVC bottom plate, a piece of absorbent filter paper being arranged at one end of the at least one surface of the PVC bottom plate and being adjacent to the nitrocellulose film, a bonding gasket being arranged at the other end of the at least one surface of the PVC bottom plate and being adjacent to the nitrocellulose film, and a sample gasket being arranged at one end of the nitrocellulose film and being adjacent to the bonding gasket. A detection line and a quality control lineare arranged at positions, being adjacent to the bonding gasket and the absorbent filter paper, of the nitrocellulose film. According to the test strip, salmonella of a plurality of serum groups canbe detected rapidly and simply in a broad spectrum manner; and thus the possibly polluted salmonella in the food can be detected conveniently and rapidly.

Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a test strip for detecting broad-spectrum Salmonella in food and methods for preparing and using the same. Background technique

[0002] Salmonella is the largest group of enteric pathogenic bacteria, mainly stored in livestock and poultry and eggs, and has the characteristics of fast reproduction and strong survival ability. The pathogenic substances of Salmonella include pili, bacterial (O) antigen, endotoxin and enterotoxin. Pili and O antigen are related to their invasiveness, endotoxin causes local intestinal inflammation and systemic poisoning symptoms, and some Salmonella can produce cholera-like enterotoxin, leading to severe diarrhea. Salmonella is contagious and can easily cause mass infections with serious consequences. It has been reported that most Salmonella infections in humans are directly or indirectly related to animal food, and among all kinds of bacterial food p...

Examples

preparation example Construction

[0038] In one embodiment, the preparation method includes the following steps:

[0039] a. utilize chloroauric acid solution and sodium citrate solution to prepare colloidal gold, obtain colloidal gold solution;

[0040] b. Under alkaline conditions, adding Salmonella polymonoclonal antibody DSP1 to the colloidal gold solution for labeling treatment, after sealing, centrifugation, and resuspension, the colloidal gold-labeled Salmonella polymonoclonal antibody DSP1 was obtained;

[0041] c. Soak the glass cellulose membrane in the conjugation pad treatment solution to prepare a gold standard pad; use colloidal gold-labeled Salmonella polymonoclonal antibody DSP1 to spray gold on the gold standard pad, and dry to obtain a conjugation pad ;

[0042] d. Take the Salmonella monoclonal antibody SP1 and the goat anti-rabbit IgG antibody, and draw them to the positions of the detection line and the quality control line of the nitrocellulose membrane with a film-drawing instrument;

...

Embodiment 1

[0066] A test strip for detecting broad-spectrum salmonella in food

[0067] Such as figure 1 As shown, test strips for detecting broad-spectrum Salmonella in food,

[0068] Including a PVC bottom plate and a nitrocellulose membrane 4 combined in the middle of at least one surface of the PVC bottom plate 1, and an absorbent filter paper 5 arranged at one end of at least one surface of the PVC bottom plate 1 and adjacent to the nitrocellulose membrane 4. The other end of at least one surface of the PVC bottom plate 1 and the bonding pad 3 adjacent to the nitrocellulose membrane 4, and the sample adjacent to the bonding pad 3 and arranged at the end away from the nitrocellulose membrane 4 Pad 2; wherein, the nitrocellulose membrane 4 is provided with a detection line 41 and a quality control line 42 near the binding pad 3 and the water-absorbing filter paper 5 respectively; the detection line 41 is coated with Salmonella monoclonal antibody SP1; the The quality control line 42...

Embodiment 2

[0070] A method for preparing a test strip for detecting broad-spectrum salmonella in food, comprising the steps of:

[0071] a. Measure 120mL of purified water into a conical flask, heat to 340°C, stir at 300rpm until boiling, add 2mL of 1% chloroauric acid solution, heat for 2min, and quickly add 2mL of 2% lemon Sodium acid solution, continue heating for 5min, the color of the liquid is stable to wine red, cool at room temperature, and obtain a colloidal gold solution with a particle size of 35-45nm;

[0072] b. Vector take 10mL colloidal gold solution, add 0.2M K 2 CO 3 solution, adjust the pH to 8.2±0.05, place it on a shaker at a speed of 80rpm for 5min, add 150μg of Salmonella antibody, carry out labeling treatment for 30min, add 100M 10% BSA solution, and carry out blocking treatment for 30min, at 15000rpm, 4°C Centrifuge for 15 minutes, discard the supernatant, and resuspend the precipitate with 600 μL gold-labeled resuspension solution to obtain colloidal gold-label...

technical field

[0001] The invention relates to the field of biotechnology, in particular to a test strip for detecting broad-spectrum Salmonella in food and methods for preparing and using the same. Background technique

[0002] Salmonella is the largest group of enteric pathogenic bacteria, mainly stored in livestock and poultry and eggs, and has the characteristics of fast reproduction and strong survival ability. The pathogenic substances of Salmonella include pili, bacterial (O) antigen, endotoxin and enterotoxin. Pili and O antigen are related to their invasiveness, endotoxin causes local intestinal inflammation and systemic poisoning symptoms, and some Salmonella can produce cholera-like enterotoxin, leading to severe diarrhea. Salmonella is contagious and can easily cause mass infections with serious consequences. It has been reported that most Salmonella infections in humans are directly or indirectly related to animal food, and among all kinds of bacterial food p...

Examples

preparation example Construction

[0038] In one embodiment, the preparation method includes the following steps:

[0039] a. utilize chloroauric acid solution and sodium citrate solution to prepare colloidal gold, obtain colloidal gold solution;

[0040] b. Under alkaline conditions, adding Salmonella polymonoclonal antibody DSP1 to the colloidal gold solution for labeling treatment, after sealing, centrifugation, and resuspension, the colloidal gold-labeled Salmonella polymonoclonal antibody DSP1 was obtained;

[0041] c. Soak the glass cellulose membrane in the conjugation pad treatment solution to prepare a gold standard pad; use colloidal gold-labeled Salmonella polymonoclonal antibody DSP1 to spray gold on the gold standard pad, and dry to obtain a conjugation pad ;

[0042] d. Take the Salmonella monoclonal antibody SP1 and the goat anti-rabbit IgG antibody, and draw them to the positions of the detection line and the quality control line of the nitrocellulose membrane with a film-drawing instrument;

...

Embodiment 1

[0066] A test strip for detecting broad-spectrum salmonella in food

[0067] Such as figure 1 As shown, test strips for detecting broad-spectrum Salmonella in food,

[0068] Including a PVC bottom plate and a nitrocellulose membrane 4 combined in the middle of at least one surface of the PVC bottom plate 1, and an absorbent filter paper 5 arranged at one end of at least one surface of the PVC bottom plate 1 and adjacent to the nitrocellulose membrane 4. The other end of at least one surface of the PVC bottom plate 1 and the bonding pad 3 adjacent to the nitrocellulose membrane 4, and the sample adjacent to the bonding pad 3 and arranged at the end away from the nitrocellulose membrane 4 Pad 2; wherein, the nitrocellulose membrane 4 is provided with a detection line 41 and a quality control line 42 near the binding pad 3 and the water-absorbing filter paper 5 respectively; the detection line 41 is coated with Salmonella monoclonal antibody SP1; the The quality control line 42...

Embodiment 2

[0070] A method for preparing a test strip for detecting broad-spectrum salmonella in food, comprising the steps of:

[0071] a. Measure 120mL of purified water into a conical flask, heat to 340°C, stir at 300rpm until boiling, add 2mL of 1% chloroauric acid solution, heat for 2min, and quickly add 2mL of 2% lemon Sodium acid solution, continue heating for 5min, the color of the liquid is stable to wine red, cool at room temperature, and obtain a colloidal gold solution with a particle size of 35-45nm;

[0072] b. Vector take 10mL colloidal gold solution, add 0.2M K 2 CO 3 solution, adjust the pH to 8.2±0.05, place it on a shaker at a speed of 80rpm for 5min, add 150μg of Salmonella antibody, carry out labeling treatment for 30min, add 100M 10% BSA solution, and carry out blocking treatment for 30min, at 15000rpm, 4°C Centrifuge for 15 minutes, discard the supernatant, and resuspend the precipitate with 600 μL gold-labeled resuspension solution to obtain colloidal gold-label...

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