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

Protein suspending chip for composite detection of multiple kinds of pathogens, its production method and detection method

A suspension chip and composite detection technology, which is applied in the direction of measuring devices, biological testing, material inspection products, etc., can solve the problems of lack of models and evaluations, and achieve the effects of simple preparation methods, wide dynamic range, and good applicability

Inactive Publication Date: 2009-08-05
CHINESE ACAD OF INSPECTION & QUARANTINE
View PDF0 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether the suspension chip can detect multiple types of pathogens at the same time, whether it is suitable for rapid and direct detection from environmental samples such as powders and liquids, and its quantitative detection ability, there is still a lack of models and evaluations.

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
  • Protein suspending chip for composite detection of multiple kinds of pathogens, its production method and detection method
  • Protein suspending chip for composite detection of multiple kinds of pathogens, its production method and detection method
  • Protein suspending chip for composite detection of multiple kinds of pathogens, its production method and detection method

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0044] 2. Preparation of samples to be tested

[0045] 1. Preparation of samples for single-component analysis

[0046] The concentration of Yersinia pestis stock solution was 10 8 cfu / mL, the concentration of anthrax spore stock solution is 10 7 cfu / mL, the stock solutions of ricin, SEB, and SARS-CoV N protein were all at 1 mg / mL. Toxin and protein samples were diluted just before use. The concentration range of the bacterial suspension was 10 1 -10 8 cfu / mL, the concentration range of spores is 10 2 -10 7 cfu / mL, toxin and protein concentrations ranged from 10 pg / mL to 5 μg / mL. The bacteria to be analyzed are diluted with PB into 10-fold different gradients, and the toxins and proteins are diluted with PB into 4-fold different gradients. The concentration of several samples is lower than the sensitivity of the detection. High-concentration samples should make the binding site of the encoded microspheres in the saturation state.

[0047] 2. Preparation of Mixed Sampl...

Embodiment 1

[0053] Embodiment 1, the preparation of protein suspension chip

[0054] A. Activation of encoded microspheres

[0055] Select 5 kinds of microspheres to label Yersinia pestis antibody (No. 028), anthrax spore antibody (No. 025), SEB antibody (043), ricin antibody (027), SARS-CoV N protein antibody (No. 044), and take 100 μL (1.25×10 6 pcs) encoded microspheres into a 1.5mL centrifuge tube, centrifuge at 14000g, carefully aspirate and discard the supernatant. Add 100 μL of microsphere washing buffer to suspend, shake and sonicate, centrifuge at 14000g, carefully aspirate and discard the supernatant. Add 100 μL of microsphere activation buffer, then add 10 μL of freshly prepared EDC (50 mg / mL), then add 10 μL of freshly prepared 50 mg / mL carboxyl-active biotin (ie Sulfo-NHS-biotin, SH-active biotin) and shake at room temperature for 20 minutes. Add 150 μL of PBS (pH7.4), shake, centrifuge at 14000 g, carefully aspirate and discard the supernatant. Add 100 μL of PBS (pH 7.4...

Embodiment 2

[0067] Embodiment 2, the improvement of the sensitivity of target pathogen and dynamic detection range

[0068] A. Preparation of samples to be tested

[0069] Yersinia pestis stock solution (10 8 cfu / mL) and anthrax spore stock solution (10 7 cfu / mL) was diluted 10 times with PB to form a series of concentration gradient samples, and ricin, SEB, and SARS-CoVN protein (stock solution 1 mg / mL) were diluted 4 times with PB to form a series of concentration gradient samples. The concentration range of Yersinia pestis suspension is 10 1 ~10 8 cfu / mL, the concentration range of anthrax spores is 10 2 ~10 7 cfu / mL, the concentrations of ricin, SEB, and SARS-CoV N protein ranged from 10 pg / mL to 5 μg / mL.

[0070] B. Testing of samples

[0071] All reactions in the detection process were carried out on a 96-well filter plate, and the detection process was as follows:

[0072] (1) Add 50 μL of working solution containing corresponding coded microspheres to each well, wash with ...

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 a method for preparing and detecting a protein suspension chip for compound detection of various pathogens. The pathogens comprise Yersinia pestis, Bacillus anthracis, staphylococcal enterotoxin B, SEB, ricin and SARS-CoV. Substantive tests prove that the method for preparing the protein suspension chip is simple, convenient and rapid and has rapid and accurate detection effect, high sensitivity, wide range of detection, strong specificity and favorable applicability for actual environment samples. And meanwhile, the invention lays a foundation for setting up a multifunctional, multi-index, high-flux and standardized requirement for the compound detection of the pathogens.

Description

technical field [0001] The invention relates to the preparation and detection method of a protein suspension chip for composite detection of various pathogens. The pathogens include bacteria, bacterial spores, bacterial toxins, plant toxins and viruses, respectively selected from Yersinia pestis , Bacillus anthracis (Bacillus anthracis), staphylococcal enterotoxin B (staphylococcal enterotoxinB, SEB), ricin (ricin) and severe acute respiratory syndrome coronavirus (SARS-CoV) as representatives. Background technique [0002] After entering the 20th century, biological threats continued unabated. The EHEC 0157:H7 epidemic in Japan in 1996, the global SARS epidemic in 2003, the anthrax mail incident in the United States in 2001, and the ricin incident in Minnesota in the United States in 1995, etc. There are countless examples of human beings facing biological threats. Man-made bioterrorism incidents may occur at any time, which arouses people's fear of biological threats. Ho...

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): G01N33/68G01N33/546G01N21/76
Inventor 王静孙肖红杨宇胡孔新张晓龙杨瑞馥
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
Features
  • Generate Ideas
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com