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Staphylococcus aureus enterotoxin detection kit and preparation and use methods thereof

The technology of a staphylococcus enterotoxin detection kit is applied in the field of immunodetection of Staphylococcus aureus enterotoxin, which can solve the problems of high specificity, long time period, weak specificity, etc. Simple operation and high sensitivity effect

Inactive Publication Date: 2016-06-08
HUAAN MAGNECH BIO TECH
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AI Technical Summary

Problems solved by technology

The results of the reverse latex agglutination test are fast, and the sensitivity of the detection of enterotoxin can also reach the nanogram level, but the existing problem is the interference of Staphylococcus protein A, which affects the accuracy of the results
The PCR method has high sensitivity and specificity, but requires specialized experimental technicians and standardized laboratories, and is not suitable for large-scale epidemiological investigations and application in primary health units
At the same time, but this method needs to sequence the results in order to finally determine the serotype of the enterotoxin, and the time period is long
③Gene probe: The isotope probe method has high specificity, strong sensitivity and good repeatability, but it is difficult to popularize due to radioactive problems
Non-radioactive probes are mainly biotin probes, which have the disadvantage of poor signal-to-noise ratio in practical applications
④Superantigen technology, the main advantage of this method is that it can directly detect the biological activity of SEA and SEB, but the specificity is not strong, and it is still in the stage of in vitro experimental research
[0005] Enzyme-linked immunosorbent assay (ELISA) has the advantages of high efficiency, sensitivity, and easy promotion at the grassroots level, and can be directly sampled for detection. It has a high consistency with HPLC and GC-MS detection. At present, there is no ELISA method for detection at home and abroad. Report on Enterotoxin SEA and SEB of Staphylococcus aureus

Method used

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Examples

Experimental program
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Effect test

specific Embodiment approach 1

[0020] Specific embodiments 1. Preparation of monoclonal antibody against Staphylococcus aureus enterotoxin SEA

[0021] 1. Acquisition of immune spleen cells

[0022] 1.1 Immunization of animals

[0023] Select 6-week-old, female, BALB / C mice, mix and emulsify the SEA antigen with Freund's complete adjuvant in equal amounts, and inject it subcutaneously at multiple points in the abdomen with an immunization dose of 50 μg for basic immunization.

[0024] Before immunization, the blood was collected by docking the tail, and the serum was separated and stored at -20°C as the serum of normal control mice. After 4 weeks, the second immunization was carried out, the dose of antigen was the same as that of the basic immunization, the antigen was mixed and emulsified with Freund's incomplete adjuvant, and the mice were boosted with immunization. Every 4 weeks, the third immunization was carried out, and the immunization conditions were the same as the second time. On the 7th to 1...

specific Embodiment approach 2

[0044] Specific embodiment two, Staphylococcus aureus enterotoxin SEB monoclonal antibody preparation

[0045] Select 6-week-old, female, BALB / C mice, mix and emulsify SEB antigen with Freund's complete adjuvant in equal amounts, and inject it subcutaneously at multiple points in the abdomen with an immunization dose of 50 μg for basic immunization. Other steps are the same as in the specific example 1 for the preparation of SEA monoclonal antibody.

specific Embodiment 3

[0046] Specific Example 3. Enzyme-labeled Staphylococcus aureus enterotoxin SEA antibody

[0047] Dissolve 4mg of HRP in 1ml of water, add 0.2ml of 1M sodium periodate to make activated HRP; use 0.5MpH10CB to adjust the pH of the activated HRP to 9.5, take 2mg of Staphylococcus aureus enterotoxin SEA monoclonal antibody, and complete the dialysis.

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Abstract

The invention discloses a staphylococcus aureus enterotoxin detection kit and preparation and use methods thereof; the kit includes staphylococcus aureus enterotoxin SEA and SEB specific antibodies and enzyme-labeled staphylococcus aureus enterotoxin SEA and SEB antibodies, has the characteristics of high specificity, high sensitivity, high precision, high accuracy and the like and can quickly detect melamine remaining in feeds and animal products.

Description

technical field [0001] The invention relates to a staphylococcus aureus enterotoxin SEA and SEB detection kit and a preparation and use method thereof, belonging to the field of immunological detection of staphylococcus aureus enterotoxin. Background technique [0002] Staphylococcus aureus is a highly pathogenic Gram-positive bacterium, and its drug resistance continues to increase, which has become one of the most important food and health safety problems in the world today. Staphylococcus aureus enterotoxin is a kind of bacterial toxin with superantigen activity secreted by Staphylococcus aureus, which can be divided into seven types according to serological methods. [0003] Staphylococcus aureus is a common bacterium in hospital infection. Staphylococcus aureus enterotoxin can cause symptoms of toxic shock and plays a key role in purulent infection. In addition, it has been found that Staphylococcus aureus enterotoxin is related to many allergies and autoimmune diseas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569
Inventor 柳家鹏王莹
Owner HUAAN MAGNECH BIO TECH
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