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Method for simultaneously detecting multiple protein markers of nutrition and health conditions

A health status and protein technology, applied in measurement devices, biological tests, material inspection products, etc., can solve the problem of poor resolution of trace detection values ​​and information capture, unsatisfactory sealing effect and sample dilution effect, and inability to multiple target proteins. Simultaneous detection and other problems to achieve the effect of reducing non-specific binding phenomenon, improving detection efficiency, and improving detection accuracy and signal stability

Pending Publication Date: 2022-03-18
NAT INST FOR NUTRITION & HEALTH CHINESE CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] It is also a technology that uses the principle of antigen-antibody specific binding to detect target proteins, but its disadvantage is that only one target protein can be detected at a time, and it is impossible to simultaneously detect multiple target proteins in one test; and its sample requirements Large, relatively low detection efficiency
[0008] It is a technology that uses immunoturbidimetric technology or chemiluminescence technology to detect target proteins, but its disadvantage is that only one target protein can be detected at a time, and it is impossible to simultaneously detect multiple target proteins in one test; and its samples need Large amount, relatively low detection efficiency
However, none of the existing detection methods can support the completion of the above work.
[0010] On the other hand, the blocking effect of the blocking solution in the existing kits that can detect other protein markers at the same time is not ideal, which leads to an abnormally high detection signal value after the implementation of the blocking link, which means that it may There is insufficient sealing, that is, non-specific signal values ​​may appear and the detection error will be increased, which will greatly reduce the detection sensitivity
[0011] Moreover, the film used for the oscillation reaction used in the existing detection kits cannot overcome the phenomenon of "splattering and hole-pouring" that occurs during the tearing process of the reaction magnetic beads adhered to the film due to oscillation, so it cannot be avoided. The appearance of abnormal detection signal value; the blocking solution and sample diluent used in the existing sample detection kits are not ideal in terms of sealing effect and sample dilution effect, resulting in distortion of the detection signal value and poor sensitivity. The ability to distinguish and capture information is poor

Method used

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  • Method for simultaneously detecting multiple protein markers of nutrition and health conditions
  • Method for simultaneously detecting multiple protein markers of nutrition and health conditions
  • Method for simultaneously detecting multiple protein markers of nutrition and health conditions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Concrete steps are as follows in the present embodiment:

[0132] Step 1: Reagent preparation, select appropriate amount of magnetic beads, cleaning solution, activation solution, PBS buffer, blocking solution, storage solution, target protein-specific antibody and biotin-labeled detection antibody for use;

[0133] Step 2: Equipment preparation, prepare the horizontal oscillator, desktop centrifuge, vortex analyzer, horizontal magnetic plate, magnetic frame, pipette, ultrasonic cleaning machine, multi-functional high-speed refrigerated centrifuge, MAGPIX flat panel detector, and adjust to standby mode;

[0134] Step 3: Activate the magnetic beads, as follows:

[0135] Step 3.1: Put the magnetic beads in the EP tube, seal the EP tube, fix the EP tube on a horizontal shaker, oscillate horizontally at 900rpm for 2min, vortex for 30s, 70HZ water bath ultrasonic for 15s, vortex for 30s, and then statically Place, take 50ul magnetic beads and put them in EP reaction tubes,...

Embodiment 2

[0170] In this example

[0171] The blocking solution consists of 50ml0.9mol / L Tris solution and 10ml1mol / L phosphate buffer, then add 10ml of 100mM NaCl, dilute to 100ml with purified water, and adjust the pH to the same as in Example 1. same.

[0172] The composition of diluent is, in 0.9% physiological saline, add phosphate (final concentration is 1mol / L), glucose (final concentration 10mmol / L), triglyceride (final concentration 2mmol / L), calf serum white protein (BSA, final concentration 0.8%).

[0173] And the condition of the oscillation treatment in step 6.12 is to shake the detection plate at 1400rpm for 30 seconds, then at 700rpm, shake for 15 minutes and then test on the machine. The rest are the same as in Example 1. The specific test results are shown in Table 5 below.

Embodiment 3

[0175] In this example

[0176] The blocking solution is composed of 45ml0.2mol / L Tris solution and 5ml1mol / L phosphate buffer, then add 5ml of 100mM NaCl, adjust the volume to 100ml with pure water, and adjust the pH to the same as in Example 1. same.

[0177] The composition of diluent is, in 0.8% physiological saline, add phosphate (final concentration is 0.1mol / L), glucose (final concentration 5mmol / L), triglyceride (final concentration 1mmol / L), calf serum Albumin (BSA, final concentration 0.5%) was prepared.

[0178] And the condition of the oscillation treatment in step 6.12 is to shake the detection plate at 1100rpm for 30 seconds, then at 700rpm, shake for 10 minutes and then test on the machine. The rest are the same as in Example 1. The specific test results are shown in Table 5 below.

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Abstract

The invention relates to a method for simultaneously detecting a plurality of protein markers for nutrition and health conditions, which comprises the following steps of: 1, reagent preparation, 2, equipment preparation, 3, magnetic bead activation, 4, antibody combination and 5, closed storage. And step 6, detecting multiple proteins in the detection plate. The method has the advantages that the protein markers capable of evaluating the nutritional status are subjected to high-throughput detection, and multiple protein markers capable of evaluating the nutritional status of the human body can be accurately detected at the same time in one-time detection, so that system errors are reduced; the systematic, comprehensive and accurate evaluation effect on the human body nutrition status is improved, and the detection efficiency is improved; the method comprises the following steps of: coating paraffin wax on a film corresponding to each hole in a film pasting oscillation link; according to the confining liquid and the sample diluent, the detection efficiency can be integrally improved.

Description

technical field [0001] The present invention relates to the field of nutritional health detection, in particular to a method for simultaneously detecting multiple protein markers of nutritional health status in the detection of multiple protein markers involved in the evaluation of human nutritional health status in the field of nutritional health. Background technique [0002] At present, the content of evaluating the basic nutritional health status of the human body must include six aspects, namely: protein nutritional status, fat nutritional status, iron nutritional status, vitamin A nutritional status, cardiovascular health status, and human tumor status. The evaluation of these six aspects of nutritional health status must be detected by different protein markers, which are: prealbumin (reflecting protein nutritional status), apolipoprotein B (reflecting human fat nutritional status), ferritin, trans Ferritin receptor and C-reactive protein (response to iron nutritional...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574G01N33/543G01N33/531
CPCG01N33/6893G01N33/57473G01N33/57476G01N33/54326G01N33/531G01N2333/47G01N2333/705G01N2333/4737G01N2333/76G01N2333/775G01N2800/60G01N2800/02G01N2800/32
Inventor 殷继永霍军生孙静刘婷婷牛江平
Owner NAT INST FOR NUTRITION & HEALTH CHINESE CENT FOR DISEASE CONTROL & PREVENTION