Search

CN-121994581-A - Immunomagnetic bead method kit for enriching staphylococcus aureus enterotoxin B from complex matrix

CN121994581ACN 121994581 ACN121994581 ACN 121994581ACN-121994581-A

Abstract

The invention provides an immunomagnetic bead kit for enriching staphylococcus aureus enterotoxin B from a complex matrix, and belongs to the technical field of food detection. A kit for enriching staphylococcus aureus enterotoxin B (SEB) comprises a carrier coupled with an anti-staphylococcus aureus enterotoxin B antibody and an enrichment solution, wherein the enrichment solution comprises 0.05M phosphate buffer solution containing 0.5% -1.2% NaCl, 0.08% -0.12% KCl and 0.08% -0.12% Tween20, and the pH value is 7.1-7.4. According to the kit, SEB in a sample to be detected is enriched under the condition of enrichment liquid by coupling the antibody of the target SEB with the magnetic beads, so that the purposes of concentrating and detecting target substances and purifying the target substances are achieved, and the detection sensitivity is effectively improved and the matrix interference is reduced.

Inventors

  • HU CHUNLI
  • WANG XI
  • HUA LING
  • GENG QIUYING
  • TIAN ZHENGAN

Assignees

  • 上海国际旅行卫生保健中心(上海海关口岸门诊部)

Dates

Publication Date
20260508
Application Date
20260211

Claims (10)

  1. 1. The kit for enriching the staphylococcus aureus enterotoxin B is characterized by comprising a carrier coupled with an anti-staphylococcus aureus enterotoxin B antibody and an enrichment solution; The enrichment solution comprises 0.05M phosphate buffer solution containing 0.5% -1.2% of NaCl, 0.08% -0.12% of KCl and 0.08% -0.12% of Tween20, and the pH value is 7.1-7.4.
  2. 2. The kit for enriching the staphylococcus aureus enterotoxin B according to claim 1, wherein the enriching liquid comprises a 0.05M phosphate buffer solution containing 0.9% of NaCl, 0.1% of KCl and 0.1% of Tween20 by mass, and the pH value is 7.2.
  3. 3. The kit for enriching a staphylococcus aureus enterotoxin B according to claim 1, wherein the carrier to which the anti-staphylococcus aureus enterotoxin B antibody is coupled comprises magnetic beads.
  4. 4. The kit for enriching a staphylococcus aureus enterotoxin B according to claim 1, wherein the anti-staphylococcus aureus enterotoxin B antibody and the carrier are coupled by an amide bond.
  5. 5. The kit for enriching a staphylococcus aureus enterotoxin B according to claim 1, further comprising a neutralization solution; The neutralization solution is 1M Tris buffer solution, and the pH value is 8.5.
  6. 6. The use of the kit for enriching staphylococcus aureus enterotoxin B according to any one of claims 1-5 in detecting the sample for enriching staphylococcus aureus enterotoxin B.
  7. 7. The use according to claim 6, wherein the enrichment temperature of the enriched staphylococcus aureus enterotoxin B is 36-38 ℃ and the enrichment time is 28-47 min.
  8. 8. The use according to claim 6, wherein the elution temperature of the enriched s.aureus enterotoxin B is 60-70 ℃.
  9. 9. The use according to any one of claims 6 to 8, wherein the test sample comprises at least one of water, food and biological fluids.
  10. 10. A kit for detecting staphylococcus aureus, comprising the kit for enriching staphylococcus aureus enterotoxin B according to any one of claims 1 to 5.

Description

Immunomagnetic bead method kit for enriching staphylococcus aureus enterotoxin B from complex matrix Technical Field The invention belongs to the technical field of food detection, and particularly relates to an immunomagnetic bead method kit for enriching staphylococcus aureus enterotoxin B from a complex matrix. Background Staphylococcus aureus (Staphylococcus aureus) is a gram-positive coccus widely existing in nature and is one of common food-borne human and animal co-occurrence pathogens. During the processing and storage of food, staphylococcus aureus is extremely easy to pollute high-protein food (such as meat, dairy products and the like), and mass reproduction and toxicity production are carried out under proper conditions, so that food poisoning is caused. The strain not only can cause skin and soft tissue infection, but also can cause enterotoxins (Staphylococcal enterotoxins, SEs) produced by the strain to be an important threat to global food safety. SEs has extremely strong heat resistance, is difficult to destroy at conventional cooking temperature, and can cause severe poisoning symptoms such as vomiting and diarrhea in human body by extremely small amount (ng). Currently identified SEs can be classified into 10 serotypes (SEA, SED, SEB, etc.) based on antigenic differences, with type B enterotoxins (SEBs) having been of interest in food safety monitoring due to their superantigenic activity (ability to non-specifically activate large numbers of T cells and initiate cytokine storms), higher thermostability (heating at 100 ℃ for 30: 30min remains active) and resistance to protease digestion (resulting in SEBs being difficult to completely eliminate during food processing). The characteristics make the establishment of an efficient and accurate SEB detection method an important link for food safety control. Currently, detection methods for SEB mainly include conventional methods such as microbial culture methods, immunoassay methods (e.g., ELISA), and molecular biology techniques (e.g., solid state fluorescent PCR). These methods, although reliable, have the disadvantage of low detection sensitivity. Disclosure of Invention The invention aims to provide an immunomagnetic bead method kit for enriching staphylococcus aureus enterotoxin B from a complex matrix, which is used for enriching SEB in a sample to be detected by coupling magnetic beads with antibodies targeting SEB, so as to achieve the purposes of concentrating detection target substances and purifying target substances, thereby effectively improving detection sensitivity and reducing matrix interference. The invention provides a kit for enriching staphylococcus aureus enterotoxin B, which comprises a carrier coupled with an anti-staphylococcus aureus enterotoxin B antibody and an enrichment solution; The enrichment solution comprises 0.05M phosphate buffer solution containing 0.5% -1.2% of NaCl, 0.08% -0.12% of KCl and 0.08% -0.12% of Tween20, and the pH value is 7.1-7.4. Preferably, the enrichment solution comprises 0.05M phosphate buffer solution containing NaCl with the mass concentration of 0.9%, KCl with the mass concentration of 0.1% and Tween20 with the mass concentration of 0.1%, and the pH value is 7.2. Preferably, the carrier to which the anti-staphylococcus aureus enterotoxin B antibody is coupled comprises magnetic beads. Preferably, the anti-staphylococcus aureus enterotoxin B antibody and the carrier are coupled by an amide bond. Preferably, the method further comprises a neutralization solution and 0.4-0.6M glycine solution; The neutralization solution comprises 1M Tris buffer with pH value of 8.5. The invention provides application of the kit for enriching staphylococcus aureus enterotoxin B in detecting sample enrichment staphylococcus aureus enterotoxin B. Preferably, the enrichment temperature of the enriched staphylococcus aureus enterotoxin B is 36-38 ℃, and the enrichment time is 28-47 min. Preferably, the elution temperature of the enriched staphylococcus aureus enterotoxin B is 60-70 ℃. Preferably, the test sample comprises at least one of water, food and biological fluids. The invention provides a kit for detecting staphylococcus aureus, which comprises the kit for enriching staphylococcus aureus enterotoxin B. The invention provides a kit for enriching staphylococcus aureus enterotoxin B, which comprises a carrier coupled with an anti-staphylococcus aureus enterotoxin antibody and an enrichment solution, wherein the enrichment solution comprises 0.05M phosphate buffer solution containing 0.5-1.2% of NaCl, 0.08-0.12% of KCl and 0.08-0.12% of Tween20 by mass, and the pH value is 7.1-7.4. According to the invention, after the anti-staphylococcus aureus enterotoxin B antibody is coupled with the carrier, the anti-staphylococcus aureus enterotoxin B antibody is utilized to capture and detect staphylococcus aureus enterotoxin B in a sample under the condition of specific enrichment liquid, and then the carrier char