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Protein G staphylococcus aureus enterotoxin kit and preparation method thereof

A staphylococcal intestinal, golden yellow technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of difficult popularization, high detection cost, low specificity, etc. Effect

Inactive Publication Date: 2016-02-24
HUAAN MAGNECH BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some of these methods have shortcomings such as low sensitivity, low specificity, detection limitations, and high detection costs, which are not easy to be popularized and used.

Method used

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  • Protein G staphylococcus aureus enterotoxin kit and preparation method thereof
  • Protein G staphylococcus aureus enterotoxin kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment 1

[0028] Specific Example 1. Establishment of the detection method of Staphylococcus aureus enterotoxin A ELISA kit

[0029] 1. Preparation of SEA antibody

[0030] Immunize Balb / c mice with Staphylococcus aureus enterotoxin A, mix the antigen and the same amount of Freund’s complete adjuvant, the immunization dose is 10μg / mouse, multiple injections on the back, abdominal cavity and sole of the plant, 30 days after the first immunization , Every 14d immunization, after immunization 6 times, take the tail blood for testing. The antibody titer in serum was determined by immunodiffusion method. Then, the antigen and the same amount of Freund's incomplete adjuvant were mixed, and the last boost before fusion was performed. Take spleen cells and myeloma cells for fusion, clone and screen to prepare anti-SEA monoclonal antibodies.

[0031] 2. Purification and identification of mAb

[0032] The mAb ascites was purified by conventional affinity chromatography, the antibody concentration was...

specific Embodiment 2

[0046] Specific Example 2. Establishment of SEBEL1SA Double Antibody Sandwich Method

[0047] 5. Production of standard curve

[0048] The standard SEA concentration is: 0, 1, 2, 4, 8, 16ppb, and the corresponding absorbance values ​​are 0.11, 0.302, 0.689, 1.208, 1.655, 1.988. The abscissa of the standard curve is the concentration of the standard product, and the ordinate is the absorbance value. , The correlation coefficient R2=0.9998.

[0049] 6. Repeatability test results of ELISA detection technology for SEA of protein G

[0050] The samples were spiked with 2ppbSEA standards, and 11 samples were extracted and tested. The results are as follows:

[0051] 1234567891011 2.052.111.981.921.932.011.961.932.011.942.06

[0052] The mean value of 11 repeated measurements was 1.99ppb, the variance (SD) was 0.06, and the coefficient of variation (CV) was 3.15%.

[0053] Specific Example 2. Establishment of the detection method of Staphylococcus aureus enterotoxin BELISA kit

[0054] Th...

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Abstract

The invention relates to a protein G staphylococcus aureus enterotoxin kit and a preparation method thereof. Protein G is adopted to preprocess an enzyme labeling plate, and quick and accurate detection on staphylococcus aureus enterotoxin A / B is realized by combining with an ELISA double-antibody sandwich method. The kit consists of the 96-hole enzyme labeling plate, a staphylococcus aureus enterotoxin A-resistant protein G recombinant protein monoclonal antibody, a staphylococcus aureus enterotoxin B-resistant monoclonal antibody, a staphylococcus aureus enterotoxin antibody enzyme labeling object containing a horseradish peroxidase label, a sample diluent, a cleaning solution and a stop buffer. The protein G staphylococcus aureus enterotoxin kit can quickly and effectively detect the residual level of tony red in a to-be-detected sample, and also has the characteristics of simple operation, good specificity, high sensitivity, and the like.

Description

technical field [0001] The invention relates to an ELISA detection kit for Staphylococcus aureus enterotoxin coated with protein G, which belongs to the field of rapid detection of Staphylococcus aureus enterotoxin. Background technique [0002] Staphylococcus aureus enterotoxins (SEs) are secreted by Staphylococcus aureus and can stimulate the central nervous system to cause poisoning reactions. The molecular weight of SEs is 26-30kDa. It is easily soluble in water and salt solution. It is stable in nature and cannot be digested by many protein enzymes. It is also very heat-resistant and cannot be destroyed by pasteurization or general cooking methods. In addition to the five most important serotypes, SEA, SEB, SEC, SED, and SEE, which have been identified for a long time, SEF, SEG, SEH, SEI, SEJ, SEK, SEL, SEM, SEN, SEO, SEP, SEQ, SER and other serotypes. Among them, food poisoning caused by SEA-SEE accounts for 95% of food poisoning caused by Staphylococcus aureus. [...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569
CPCG01N33/577G01N33/56911
Inventor 王莹张彦明
Owner HUAAN MAGNECH BIO TECH
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