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

Kit for detecting procalcitonin in blood and preparation method

A technology of procalcitonin and kits, applied in the direction of biological testing, material inspection products, etc., to achieve the effects of reducing non-specific adsorption, low background and high, and enhancing immune signals

Active Publication Date: 2019-11-12
NINGBO AUCHEER BIOTECHNOLOGY CO LTD
View PDF12 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the test results can only meet the requirements of semi-quantitative

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A kit for detecting procalcitonin in blood, comprising a quartz needle coated with a capture antibody, a biotin-labeled detection antibody, fluorescein-labeled streptavidin, and a washing buffer.

[0021] The polysaccharide complex is coated on the solid phase carrier, and the polysaccharide complex is a deionized aqueous solution containing 0.5-2.5% chitosan, 0.1-0.5% glycerin and 0.5-1.5% polyethylene glycol 1000 in mass fraction.

[0022] Described washing buffer is the PBS damping fluid (0.01M, pH7) containing the NaCl of 0.15~0.35M, the triton X-100 of 0.05~0.1%vol, the sodium lauryl sarcosine of 3~6g / L. .4).

Embodiment 2

[0024] A preparation method of a test kit for detecting procalcitonin in blood, comprising the steps of:

[0025] (1) Solid phase carrier coated with capture antibody: soak the lower end of the quartz needle in the polysaccharide complex for 1-3 minutes, freeze-dry at 4°C, and dilute the procalcitonin capture antibody to 1mg with 0.01M PBS buffer / mL, place the lower end of the dried quartz needle in procalcitonin capture antibody solution, react for 2 hours at 24°C for labeling, wash 3 times with 0.1M PBS buffer after labeling, and then place in blocking solution Blocking for 8-12 hours, the blocking solution is Tris-Hcl buffer containing 1% BSA, 0.1% casein, and 3% sucrose to prepare a solid-phase carrier coated with capture antibodies;

[0026] (2) Biotin-labeled detection antibody: Dilute the procalcitonin detection antibody to 1 mg / mL with 0.01M PBS buffer, and add biotin according to the mass ratio antibody:biotin=1:3-1:8 , react at 24°C for 8-12 hours for labeling, dia...

Embodiment 3

[0029] How to use the kit: put the solid-phase carrier coated with the capture antibody into the sample, take it out after 10-60s and wash it with washing buffer for 3-5 times, then put it into the biotin-labeled detection antibody for 10-60s Take it out and wash it 3-5 times with washing buffer, put it into fluorescein-labeled streptavidin for 10-60s after washing, take it out and wash it 3-5 times with washing buffer, and then use the fluorescence analysis detector to detect.

[0030]The target antigen in the sample of the present invention first binds to the capture antibody on the surface of the solid-phase carrier, and the solid-phase carrier is subsequently immersed in the biotin-labeled detection antibody and fluorescein-labeled streptavidin to carry out the binding reaction. The solid-phase carrier The surface is coated with a biocompatible polysaccharide complex, which can effectively resist non-specific binding, and the amplification of the signal is achieved by formi...

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 discloses a kit for detecting procalcitonin in blood and a preparation method. The kit comprises a solid-phase carrier coated with a captured antibody, a biotin-labeled detection antibody, fluorescein labeled streptavidin, a washing buffer solution and the like. The surface of a quartz needle is coated with a polysaccharide compound with biocompatibility, wherein the polysaccharide compound can effectively enhance the specific immune signal, and the amplification of a detection signal is realized by forming a plurality of fluorescein-streptavidin molecular layers on the surface of the polysaccharide compound; and according to the kit, SDS with relatively high washing capability and Triton-X100 are added in the washing buffer solution, and the ionic strength of the washing solution is improved, so that the non-specific interaction between the molecules is effectively controlled, the low background and the high signal-to-noise ratio of the detection result are further realized, and the sensitivity and accuracy of detection are improved.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis, in particular to a kit and a detection method for detecting procalcitonin in blood. Background technique [0002] PCT (procalcitonin, procalcitonin) is a protein derived from a single-copy gene located on chromosome 11 (11p15,4), which consists of 2800 base pairs and contains 6 exons and 5 introns. After transcription, it is translated into procalcitonin precursor in the rough endoplasmic reticulum of parafollicular cells of the thyroid, including 84 N-terminal amino acids, active calcitonin and calcitonin. The procalcitonin precursor cuts off the single sequence of nPro-CT end under the action of endogenous polypeptide enzymes to generate 116 amino acid PCT with a molecular weight of about 13kD. PCT and calcitonin have the same sequence of 32 amino acids (60~ 91 bits). [0003] PCT is the hormonally inactive calcitonin propeptide substance. The content in healthy people is very small, and i...

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
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 周义正唐静陈星星黄丹娣余城滨
Owner NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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