Immunoturbidimetric kit for detecting procalcitonin

A procalcitonin and kit technology, applied in the field of medical immunization, can solve the problems of complicated operation, narrow linear range, and increase the pain of subjects, and achieve the effects of high detection sensitivity, broadened linear range, and wide linear range

Inactive Publication Date: 2017-11-10
SONOSCAPE MEDICAL CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Radioimmunoassay can detect serum PCT in normal people, but it cannot distinguish between free PCT, bound PCT and calcitonin gene-related peptide precursor, and it takes too long (19-22h). There are certain limitations; high-performance liquid chromatography is time-consuming and difficult to automate; enzyme-linked immunosorbent assay is complicated to operate and takes a long time; it is generally qualitative or semi-quantitative, and the results have large deviations; chemiluminescence has high sensitivity It can meet the clinical PCT detection requirements, but it needs to be matched with special equipment, which is expensive; colloidal gold colorimetry is a qualitative detection, fast and easy, and can obtain the PCT concentration range, but cannot obtain accurate values
Transmission immunoturbidimetr

Method used

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  • Immunoturbidimetric kit for detecting procalcitonin
  • Immunoturbidimetric kit for detecting procalcitonin
  • Immunoturbidimetric kit for detecting procalcitonin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 kit of the present invention

[0034] 1) Preparation of reagent R1:

[0035] Weigh 24.2g Trizma-base, 7.9g NaCL, 10g BSA, 10g PEG6000, 3g SDS, 5g Tween 80, 1g EDTA and 1g sodium azide according to the weight of the formula and dissolve them in 0.8L distilled water, adjust the pH value to 8.0, set Fill to the 1L scale line, filter to get R1;

[0036] 2) Preparation of reagent R2:

[0037] Step 1: Combine carboxylated polystyrene latex microspheres with particle sizes of 80nm and 198nm in a certain proportion, wash with distilled water 3 times, add 200mmol / L PBS buffer to dilute the polystyrene latex microspheres to 0.01mg / ml;

[0038] Step 2: Add 4 mg EDC and 2 mg NHS to 10 ml of diluted latex microspheres, stir continuously at room temperature for 30 minutes, then centrifuge, wash with PBS buffer to remove unreacted EDC and NHS, and dilute to 0.01 mg / ml;

[0039] Step 3: Dissolving and diluting the PCT antibody to 1 mg / ml with Tris buffer solution of Ph8...

Embodiment 2

[0041] Embodiment 2 kit of the present invention

[0042] 1) Preparation of reagent R1:

[0043] Weigh 12.1g Trizma-base, 15g NaCL, 10g BSA, 20g PEG6000, 5g SDS, 3g Tween 80, 1g EDTA and 1g sodium azide in 0.8L distilled water, adjust the pH value to 8.0, and constant volume To the 1L scale line, filter to get R1;

[0044] 2) Preparation of reagent R2:

[0045] Step 1: Combine carboxylated polystyrene latex microspheres with particle sizes of 123nm and 220nm in a certain proportion, wash with distilled water for 3 times, add 100mmol / L PBS buffer to dilute the polystyrene latex microspheres to 0.01mg / ml;

[0046] Step 2: Add 2 mg EDC and 4 mg NHS to 10 ml of diluted latex microspheres, stir continuously at room temperature for 30 minutes, then centrifuge, wash with PBS buffer to remove unreacted EDC and NHS, and dilute to 0.01 mg / ml;

[0047] Step 3: Dissolving and diluting the PCT antibody to 1 mg / ml with Tris buffer solution of Ph8.0, then adding the PCT antibody diluent...

Embodiment 3

[0049] Embodiment 3 kit of the present invention

[0050] 1) Preparation of reagent R1:

[0051] Weigh 23.83g Hepes, 20g NaCL, 30g BSA, 30g PEG6000, 5g dodecyltrimethylammonium chloride, 3g Tween 80, 1g EDTA and 1g sodium azide according to the formula quality and dissolve them in 0.8L distilled water, adjust When the pH value reaches 8.0, set the volume to the 1L mark, and filter to obtain R1;

[0052] 2) Preparation of reagent R2:

[0053] Step 1: Combine carboxylated polystyrene latex microspheres with particle sizes of 107nm and 309nm in a certain proportion, wash with distilled water for 3 times, add 100mmol / L MES buffer to dilute the polystyrene latex microspheres to 0.01mg / ml;

[0054] Step 2: Add 2mgEDC and 4mgNHS to 10ml of diluted latex microspheres, stir continuously at room temperature for 30 minutes, then centrifuge, wash with 200mmol / LPBS buffer to remove unreacted EDC and NHS, and dilute to 0.01mg / ml;

[0055] Step 3: Dissolve and dilute the PCT antibody ...

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Abstract

The present invention relates to the field of medical immunology, particularly to an immunoturbidimetric kit for detecting procalcitonin. The kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 comprises a buffer liquid, a stabilizer, a protection agent, a coagulant, a hemolytic agent and a preservative, the reagent R2 comprises a buffer liquid, a stabilizer, a protection agent, a preservative and a polystyrene latex microsphere-procalcitonin antibody complex, and the polystyrene latex microspheres comprise two types of microspheres such as microspheres having a particle size of 80-150 nm and microspheres having a particle size of 150-400 nm. According to the present invention, the test results show that the kit has advantages of good stability and long storage period, can directly detect the PCT in whole blood, and further has advantages of high detection result accuracy, high sensitivity and wide linear detection range.

Description

technical field [0001] The invention relates to the field of medical immunity, in particular to an immunoturbidimetric detection kit for detecting procalcitonin. Background technique [0002] Procalcitonin (PCT) is a glycoprotein consisting of 116 amino acids with a molecular weight of about 13Kd. Its structure is stable and not affected by hormone activity in the body. Its half-life in the human body is 25-30h, stable Good is better. PCT is mainly produced by the liver, and the neuroendocrine cells of the spleen, kidney and lung are also important places for its production. [0003] PCT is a clinical marker and an important parameter in the diagnosis and monitoring of infections in bacterial inflammatory diseases. Widely used in hematology oncology, anesthesiology, internal medicine, transplant surgery, neonatology and pediatrics and other diagnostic and treatment departments. Under normal circumstances, PCT exists in human serum in a free form, and its level is very low...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543G01N33/531G01N21/51
CPCG01N21/51G01N33/531G01N33/54313G01N33/68
Inventor 蒋玉林刘兵陈彬陈智峰
Owner SONOSCAPE MEDICAL CORP
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