Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Amanita amanita peptide detection method for non-disease diagnosis purpose

A detection method, the technology of amanitin, which is applied in the detection field, can solve the problems of short detection window period and low detection rate of amanitin, so as to extend the detection window period, meet the sensitivity and detection limit, and be easy to promote The effect of using

Pending Publication Date: 2022-08-02
中国疾病预防控制中心职业卫生与中毒控制所
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Therefore, the technical problem to be solved by the present invention is to overcome the defects of short detection window period and low detection rate of amanitin existing in the existing amanitin detection method, thereby providing a kind of amanita for non-disease diagnosis purpose. Ointment detection method

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Amanita amanita peptide detection method for non-disease diagnosis purpose
  • Amanita amanita peptide detection method for non-disease diagnosis purpose
  • Amanita amanita peptide detection method for non-disease diagnosis purpose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The detection method of protein-bound α-amanita peptide in plasma of exposed mice includes the following operations:

[0049] (1) Preparation of samples to be tested

[0050] Take 200 μl of the plasma sample (sample to be tested) of the infected mice in a 1.5 ml centrifuge tube, add 20 μl of 1wt% trypsin solution (prepared by dissolving the solid trypsin in phosphate buffer solution) to make the final concentration of trypsin 0.11 wt%, mixed by vortex, and hydrolyzed in a water bath at 37°C for 8h. Take the solid-phase extraction column HLB, activate it with 1 mL of methanol and 1 mL of water in turn, let it stand for equilibrium, take the hydrolyzed plasma sample (220 μl in volume) and load it, and then use 1 mL of 5% methanol aqueous solution to wash, rinse After washing, 2 mL methanol was used for stepwise elution, and the eluate was collected and evaporated to dryness by vacuum centrifugation. After evaporating the solvent to dryness, add 10 μL of internal standar...

experiment example 1

[0083] This experimental example is used to verify the effect of protease hydrolysis on the detection of free α-amanita peptide in the sample:

[0084] Add α-amanita to the blank plasma of mice to prepare a spiked plasma (sample) containing 10 μg / L α-amanita peptide, then take 200 μl of the spiked plasma and add 20 μl of 0.25% trypsin solution , after mixing, hydrolyzed in water bath at 37°C for 0-12h. Take the plasma samples at different hydrolysis time points, prepare the samples to be tested according to the solid phase extraction method in the operation (1) of Example 1, and detect according to the UPLC-MS / MS method of the operation (3) in Example 1. Two samples were tested in parallel at the hydrolysis time point, and the test results are shown in Table 6. It can be seen from Table 6 that the addition of protease and the hydrolysis time within 12h have little effect on the detection results of free α-amanita peptide content in plasma.

[0085] Table 6 Effect of protease...

experiment example 2

[0088] This experimental example is used to verify the presence of protein-bound α-amanita peptide in plasma samples:

[0089]ICR male mice were injected with α-amanita toxin by intraperitoneal injection at doses of 0.33mg / kg and 0.66mg / kg, respectively, and the mice were collected at different time points within 0.5h-14d after exposure. Plasma, the method of embodiment 1 (protease hydrolysis) and comparative example 1 (unhydrolyzed) was used to detect the α-amanita peptide content in the mouse plasma collected at each time point, and the detection rate was counted. The results are shown in Table 7 shown (LOD = 0.3 μg / L).

[0090] Table 7 Detection results of different detection methods on α-amanita peptide in the plasma of the same infected mice

[0091]

[0092] As can be seen from Table 7, after the mice were exposed to α-amanita for 2d, when the method of Comparative Example 1 (not hydrolyzed) was used for detection, α-amanita could not be detected in the serum of the ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the technical field of detection, in particular to a non-disease-diagnosis amanita peptide detection method. The detection method comprises the operation of detecting amatoxin in a to-be-detected sample, wherein the to-be-detected sample is prepared by performing protein degradation treatment on the to-be-detected sample. A to-be-detected sample used in the detection method is prepared after the to-be-detected sample is subjected to protein degradation treatment, and protein in the protein binding state amatoxin can be degraded through the protein degradation treatment, so that more free state amatoxin is released. Therefore, the free amatoxin contained in the used sample to be detected is composed of the original free amatoxin and the free amatoxin released by the protein binding state amatoxin, that is, the detection method disclosed by the invention can be used for simultaneously detecting the free amatoxin and the protein binding state amatoxin contained in the sample to be detected; therefore, the detection window period of the amatoxin can be obviously prolonged, and the detection rate of the amatoxin is effectively increased.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a non-diagnostic purpose amanita peptide detection method. Background technique [0002] Mushroom poisoning is one of the main causes of death from food poisoning and one of the prominent public health problems affecting public health. According to the survey data of mushroom poisoning deaths, the main cause of death from mushroom poisoning is that poisonous mushrooms contain amanitamin. The poisoning death was caused by acute liver failure of the poisoned person caused by the amanita venom peptide contained in the mushroom. Among the known amanita toxins, α-amanita is the most toxic and the most representative amanita toxin. Therefore, research on the toxicity and chemical properties of α-amanita peptide is of great value for solving the clinical problem of mushroom amanita poisoning. [0003] "Expert Consensus on Clinical Diagnosis and Treatment of Mushroom Poisoning Conta...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N30/02G01N30/06G01N30/72G01N30/86
CPCG01N30/02G01N30/06G01N30/72G01N30/8679G01N2030/065Y02A90/10
Inventor 吴智君孙承业樊晶光李海蛟张宏顺丁春光代静郑敏程娟赵文锦
Owner 中国疾病预防控制中心职业卫生与中毒控制所
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com