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Bacillus cereus immune colloidal gold rapid detection test strip and preparation method thereof

A technology of Bacillus cereus and detection test paper, which is applied in the field of microbial detection, can solve the problems of rapid detection of Bacillus cereus and achieve high sensitivity

Pending Publication Date: 2021-09-03
UNIV OF SHANGHAI FOR SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, my country currently does not have products that can be used for rapid detection of Bacillus cereus

Method used

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  • Bacillus cereus immune colloidal gold rapid detection test strip and preparation method thereof
  • Bacillus cereus immune colloidal gold rapid detection test strip and preparation method thereof
  • Bacillus cereus immune colloidal gold rapid detection test strip and preparation method thereof

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preparation example Construction

[0036] A preparation method for bacillus cereus immune colloidal gold rapid detection test strip, comprising the following steps:

[0037] S1, carry out the preparation of colloidal gold;

[0038] S2. Determine the optimal pH of the colloidal gold-labeled antibody;

[0039] S3, determining the optimal binding amount of the colloidal gold-coupled antibody;

[0040] S4, labeling the antibody with colloidal gold;

[0041] Further, the step S1 includes:

[0042] S11. Take the chloroauric acid powder and add it to sterilized double distilled water to form a chloroauric acid solution;

[0043] S12. Take the dihydrate trisodium citrate, dissolve it in distilled water, and make a trisodium citrate solution, which is prepared and used immediately;

[0044] S13. In a flask of double-distilled water, adjust the temperature of the magnetic stirrer to 150°C, the stirring rate is 500rpm, heat to boiling, immediately add chloroauric acid, boil for 15s, stir fully and quickly add 1% triso...

Embodiment 1

[0056] The preparation of embodiment 1 bacillus cereus monoclonal antibody

[0057] 1. Preparation of Immunogen

[0058] Cultivate Bacillus cereus in LB broth for 18-24h, then perform PCR identification, wash the bacterial solution three times with sterilized PBS, and adjust its concentration to make the OD 600nm is 0.6.

[0059] 2. Preparation process of monoclonal antibody

[0060] 1) Experimental animals: Three female BALB / C mice aged 6-8 weeks, weighing about 25 g, were selected as experimental animals.

[0061] 2) Immunization method: each mouse was intraperitoneally injected with 0.4ml of immunogen, and the same dose was boosted once every 2 weeks.

[0062] 3) Blood collection: Blood was collected from the tail vein after 3 times of immunization, and the antiserum titer was determined by indirect non-competitive enzyme-linked immunoassay. After the titer no longer increased, the same amount of immunogen was injected intraperitoneally, and cell fusion was carried out ...

Embodiment 2

[0082] The preparation of embodiment 2 Bacillus cereus immune colloidal gold test strip

[0083] 1. Preparation process of colloidal gold

[0084] 1) Preparation of 1% chloroauric acid: Take 1g of chloroauric acid powder, add 100mL of sterilized double distilled water to prepare 1% chloroauric acid solution, put it at 4°C for later use.

[0085] 2) Preparation of 1% trisodium citrate: Take 0.5679 g of trisodium citrate dihydrate, dissolve it in 50 mL of double-distilled water, and prepare a 1% trisodium citrate solution, which is prepared and used immediately.

[0086] 3) Prepare colloidal gold particles by the citrate reduction method. In a 198mL flask of double distilled water, adjust the temperature of the magnetic stirrer to 150°C, the stirring rate is 500rpm, heat to boiling (about 100°C), and immediately add 1% gold chloride Add 2mL of acid, boil for 15s, stir well and quickly add 3mL of 1% trisodium citrate, time for 10min, stop heating, cool to room temperature, store...

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Abstract

The invention discloses a bacillus cereus immune colloidal gold rapid detection test strip. The test strip comprises a PVC bottom plate, a sample pad arranged on the PVC bottom plate, a combination pad adjacent to the sample pad and a water absorption pad adjacent to the combination pad, and a nitrocellulose membrane layer is arranged between the conjugate pad and the PVC bottom plate, and a detection line and a quality control line are arranged on the conjugate pad. According to the invention, the method can be used for detecting the immunogen of the bacillus cereus, and the detection sensitivity is high.

Description

technical field [0001] The invention relates to the technical field of microorganism detection, in particular to a test strip for quick detection of bacillus cereus immune colloidal gold. Background technique [0002] Bacillus cereus is a Gram-positive bacterium that causes disease in humans and animals. According to the data analysis of the National Foodborne Disease Surveillance Network, dozens of Bacillus cereus food poisoning incidents occur in my country every year, accounting for about 11.4% of the total number of bacterial food poisoning incidents, causing hundreds of people to fall ill, and this number may be far lower The actual number of poisoned people. Bacillus cereus infection can cause nausea, vomiting, abdominal pain, diarrhea, etc., and severe cases can cause electrolyte imbalance and bloody diarrhea. The pathogen mainly exists in rice products, meat products, dairy products and other finished products and their semi-finished products. In the production, pr...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/577G01N33/58G01N33/558G01N33/532B22F9/24B22F1/00B82Y40/00B82Y30/00
CPCG01N33/56911G01N33/577G01N33/587G01N33/558G01N33/532B22F9/24B82Y40/00B82Y30/00G01N2333/32G01N2469/10
Inventor 王英林张闻桐刘程方水琴田亚晨吴美娇吴有雪刘涛杨昊李斌吴娅芳姜嘉烨黄笑天刘箐
Owner UNIV OF SHANGHAI FOR SCI & TECH
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