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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
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  • 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

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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...

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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

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Application Information

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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
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