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A chemiluminescence quantitative detection kit for procalcitonin and its preparation method and detection method

A technique for quantitative detection of procalcitonin, applied in the field of immunoassay, can solve the problems of low sensitivity of the kit, narrow detection range, poor specificity of the kit, etc., and achieves simple and time-saving operation, wide detection range and good accuracy. Effect

Active Publication Date: 2016-05-11
SUZHOU HAOOUBO BIOPHARML
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the sensitivity of the kit in this patent is still low (0.018ng / ml), the specificity of the kit is also poor, and the detection range is narrow

Method used

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  • A chemiluminescence quantitative detection kit for procalcitonin and its preparation method and detection method
  • A chemiluminescence quantitative detection kit for procalcitonin and its preparation method and detection method
  • A chemiluminescence quantitative detection kit for procalcitonin and its preparation method and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: the preparation of kit

[0039] (1) Preparation of magnetic separation reagents:

[0040] Materials and Instruments:

[0041] 1. Magnetic particles: the surface contains carboxyl (COOH) active groups, and the carboxyl group content per gram of magnetic particles (dry weight) is 0.4 mmol, with superparamagnetism and a diameter of 1 μm.

[0042] 2. Streptavidin was purchased from Sigma.

[0043] 3. 2-Morpholineethanesulfonic acid (MES), carbodiimide (EDC), trishydroxymethylaminomethane (TRIS) and other reagents are all chemically pure.

[0044] 4. Analytical balance

[0045] Steps:

[0046] 1. Take 100mg of magnetic particles, magnetically separate to remove the supernatant, and resuspend in 10ml of 0.05mol / L, pH4.7 MES buffer;

[0047] 2. Add 3 mg of streptavidin and suspend at room temperature for 50 minutes;

[0048] 3. Add 0.7ml freshly prepared 10mg / ml EDC aqueous solution and suspend at room temperature for 8h;

[0049] 4. Magnetic separation, r...

Embodiment 2

[0081] Embodiment 2: the implementation of detection and the evaluation of detection effect

[0082] Materials and Instruments:

[0083] 1. The kit prepared by Example 1;

[0084] 2. The luminescent substrate and cleaning solution are produced by Suzhou Hao Oubo Bio-Pharmaceutical Co., Ltd.

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Abstract

The invention relates to a chemiluminescence quantitative detection kit for procalcitonin, and a preparation method and detection method thereof. The kit comprises a procalcitonin series standard substance, a magnetic separation reagent (magnetic particle suspension coupled with streptavidin), a first reagent (anti-procalcitonin monoclonal antibody solution containing biotin N-hydroxysuccinimide ester label) and a second reagent (anti-procalcitonin monoclonal antibody solution containing alkaline phosphatase label). The sensitivity of the kit prepared from the magnetic particle suspension coupled with streptavidin, anti-procalcitonin monoclonal antibody solution containing biotin N-hydroxysuccinimide ester label and anti-procalcitonin monoclonal antibody solution containing alkaline phosphatase label is up to 0.008ng / ml; and the kit has the advantages of high accuracy, high precision, no need of prediluting the sample, and wide detection range, and is simple and time-saving to operate.

Description

technical field [0001] The invention relates to the field of immune analysis, in particular to a chemiluminescence quantitative detection kit for procalcitonin, a preparation method and a detection method thereof. Background technique [0002] Procalcitonin (PCT) is derived from a single-copy gene located on chromosome 11 (11p15,4), which consists of 280 base pairs with 6 exons and 5 introns. After transcription, it is translated into Preprocalcitonin 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 32 amino acid sequence (60~ 91 bits). [0003] PCT is expressed by neuroendocrine cells (including C cells in thyroid, lung, and pancreatic tissues) and is cle...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N21/76
CPCG01N21/76G01N33/54326G01N33/577
Inventor 徐顺澍李永红叶兴旺周超王秀伟李庆春
Owner SUZHOU HAOOUBO BIOPHARML
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