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Kit for detecting procalcitonin with spatial proximity chemiluminescence method and a detection method of kit

A technology for detecting kits and procalcitonin, which is applied in chemiluminescence/bioluminescence, analysis by making materials react chemically, and measurement devices. It is easy to scale up production and realize the effect of full automation

Inactive Publication Date: 2019-01-04
无锡壹闪生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The latest definition of sepsis is fatal organ dysfunction caused by dysregulation of the host response to infection. reaction and other aspects, but its specific mechanism has not been fully elucidated

Method used

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Examples

Experimental program
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Effect test

preparation example Construction

[0025] 1. Preparation of calibrator

[0026] (1) Preparation of calibration product dilution: Weigh 14.1 g of potassium dihydrogen phosphate, sodium dihydrogen phosphate (2H 2 O) 3.0g, add ultrapure water to dissolve, Proclin-300 0.5 ~ 1mL, after mixing, add ultrapure water to make up to 1000mL, obtain the calibration product dilution, store at 2 ~ 8 ℃ for later use.

[0027] (2) Preparation of calibrator: the concentration of the calibrator is 0, 10ng / mL, 50ng / mL, 500ng / mL, 1000ng / mL, 10000ng / mL, dilute the purified procalcitonin to the corresponding concentration with the calibrator diluent, Store at 2-8°C for later use.

[0028] 2. Preparation of enzyme markers

[0029] (1) Weigh 5 mg of HRP and dissolve it in 1 mL of distilled water, add 0.2 mL of freshly prepared 0.1M sodium periodate solution to the supernatant, stir at room temperature in the dark for 20 min, put the above solution into a dialysis bag, and adjust to 1 mM pH 4.4 Dialyzed against sodium acetate buffer ...

Embodiment 1

[0047] This embodiment provides a detection kit for procalcitonin, including: an enzyme marker, a luminescent marker, an auxiliary agent, a trigger, and a calibrator; wherein, the calibrator includes no less than 6 different concentrations of PCT antigen calibration products and 0.1M phosphate buffer; enzyme labels include peroxidase-labeled PCT detection antibody and 0.05M phosphate buffer; luminescent labels include 9,10-dihydroacridine-labeled PCT capture antibody and 0.05M Tris buffer; the components of the auxiliary agent include luminescent auxiliary agent and pH 6.0 citrate buffer; the trigger agent is selected from 0.05M pH 8.0 Tris-HCl buffer.

[0048] 1. Preparation of calibrator

[0049] (1) Preparation of calibration product dilution: Weigh 14.1 g of potassium dihydrogen phosphate, sodium dihydrogen phosphate (2H 2 O) 3.0g, add ultrapure water to dissolve, Proclin-300 0.8mL, after mixing, add ultrapure water to make up to 1000mL to obtain the calibration product d...

Embodiment 2

[0064] This embodiment provides a method for detecting procalcitonin using a procalcitonin detection kit in a spatially adjacent chemiluminescence method, including steps:

[0065] S1: Add 25 μL of standard, 25 μL of peroxidase-labeled PCT detection antibody and 25 μL of luminescent marker to the chemiluminescence reaction tube.

[0066] S2: React in a 37°C incubator for 15 minutes.

[0067] S3: Add 5 μL of auxiliary agent to the chemiluminescent reaction tube, shake and mix, and let it stand for 1-2 minutes; then add 75 μL of trigger agent, shake and mix, detect immediately, and read the signal value.

[0068] S4: Carry out logistic four-parameter fitting on the concentration and luminescence value of the calibrator, and calculate the sample concentration through the luminescence value of the sample.

[0069] The invention adopts the spatial proximity chemiluminescence method to detect the procalcitonin, and the sensitivity can reach 0.1ng / mL. Elevated serum PCT has a speci...

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Abstract

The invention relates to a kit for detecting procalcitonin with a spatial proximity chemiluminescence method and a detection method of the kit. The kit comprises an enzyme marker, a luminescent marker, an adjuvant, a trigger and a calibrator, wherein the calibrator contains calibration products of PCT antigens with different concentrations and a 0.1 M phosphate diluent; the enzyme marker containscomponents of a peroxidase labeled PCT detection antibody and a 0.05 M phosphate buffer; the luminescent marker contains components of a 9,10-dihydracridine labeled PCT capture antibody and 0.05 M Tris buffer; the adjuvant contains components of a luminescent adjuvant and a citrate buffer; the trigger is a 0.05 M Tris buffer. The detection method as a true homogeneous chemiluminescence technologydoes not need a carrier, coating and washing processes are omitted, and the kit comprises few components, has high sensitivity and good repeatability and is easy to operate.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting procalcitonin by a spatially adjacent chemiluminescence method and a detection method thereof. Background technique [0002] Procalcitonin (PCT) is composed of amino acids and belongs to calcitonin propeptide substances without hormone activity. It has good stability and a half-life of 25 to 30 hours. PCT is generally produced in thyroxine cells, and calcitonin is formed by specific proteolysis. Under pathological conditions, PCT is stimulated and induced by inflammatory cytokines and bacterial toxins, and its secretion is produced in extra-thyroxine tissues and organs. Its levels have been reported to generally not rise in response to non-infectious inflammation. A large number of scholars agree that the level of PCT rises during bacterial infection and remains low during local inflammatory response and viral infection. This change is stable and rapid, and it i...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/577G01N33/535
CPCG01N21/76G01N33/535G01N33/577G01N2333/585
Inventor 奚伟红张平廖鸳鸯朱丹丹
Owner 无锡壹闪生物科技有限公司
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