Kit for detecting procalcitonin by spatial proximity chemiluminescence method and detection method and application thereof

A detection kit, procalcitonin technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc. Achieve fully automated, easy-to-scale production results

Pending Publication Date: 2021-08-10
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

[0005] 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
Comparison scheme
Effect test

preparation example Construction

[0026] 1. Preparation of calibrators

[0027] (1) Preparation of calibrator dilution: Weigh 14.1 g of potassium dihydrogen phosphate, sodium dihydrogen phosphate (2H 2 O) 3.0g, add ultrapure water to dissolve, Proclin-3000.5~1mL, after mixing, add ultrapure water to dilute to 1000mL to obtain calibrator dilution, store at 2~8℃ for later use.

[0028] (2) Prepare the calibrator: the concentration of the calibrator is 0, 10ng / mL, 50ng / mL, 500ng / mL, 1000ng / mL, 10000ng / mL, and the purified procalcitonin is diluted to the corresponding concentration with the calibrator diluent, Store at 2-8°C for later use.

[0029] 2. Preparation of Enzyme Labels

[0030] (1) Weigh 5mg of HRP and dissolve it in 1mL of distilled water, add 0.2mL of newly prepared 0.1M sodium periodate solution to the supernatant, and stir for 20min in the dark at room temperature. of sodium acetate buffer, overnight at 4°C.

[0031](2) Add 20 μL of 0.2M pH 9.5 carbonate buffer, then immediately add 1 mg of anti...

Embodiment 1

[0048] This embodiment provides a detection kit for procalcitonin, including: an enzyme marker, a luminescent marker, an adjuvant, a trigger, and a calibrator; wherein the calibrator includes not less than 6 calibrations of PCT antigens with different concentrations 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 solution; the components of the auxiliary agent include luminescence auxiliary agent and pH 6.0 citrate buffer; the trigger agent is 0.05M pH 8.0 Tris-HCl buffer.

[0049] 1. Preparation of calibrators

[0050] (1) Preparation of calibrator dilution: Weigh 14.1 g of potassium dihydrogen phosphate, sodium dihydrogen phosphate (2H 2 O) 3.0g, add ultrapure water to dissolve, Proclin-3000.8mL, after mixing, add ultrapure water to dilute to 1000mL to obtain calibrator dilution, store at 4°C for later use....

Embodiment 2

[0065] This embodiment provides a method for detecting procalcitonin using a procalcitonin detection kit in a spatial proximity chemiluminescence method, comprising the steps of:

[0066] 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, respectively.

[0067] S2: React in a 37°C incubator for 15 minutes.

[0068] S3: Add 5 μL of adjuvant to the chemiluminescence reaction tube, shake and mix, and let stand for 1-2 minutes; then add 75 μL of triggering agent, shake and mix, immediately detect, and read the signal value.

[0069] S4: Perform logistic four-parameter fitting on the concentration and luminescence value of the calibrator, and calculate the sample concentration through the sample luminescence value.

[0070] The invention adopts the spatial proximity chemiluminescence method to detect procalcitonin, and the sensitivity can reach 0.1ng / mL. Elevated serum PCT has a sp...

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PUM

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Abstract

The invention relates to a kit for detecting procalcitonin by a spatial proximity chemiluminescence method and a detection method and application thereof. The kit comprises an enzyme marker, a luminous marker, an auxiliary agent, a triggering agent and a calibrator, wherein the enzyme marker is prepared from the following raw material components, a peroxidase-labeled PCT detection antibody and a 0.05 M phosphate buffer solution; the luminous marker is prepared from the following raw material components, 9, 10-dihydroacridine labeled PCT (Procalcitonin) capture antibody and 0.05 MTris buffer solution; the calibrator comprises calibrators of PCT antigens with different concentrations and a 0.1 M calibrator diluent; the auxiliary agent comprises a luminous auxiliary agent and a citrate buffer solution; the 0.05 MTris buffer solution is selected as a triggering agent. The detection method for the non-treatment purpose, provided by the invention, is used as a real homogeneous chemiluminescence technology, does not need a carrier, does not have coating and washing processes, has few components in the kit, and is high in sensitivity, good in repeatability and simple to operate.

Description

[0001] This application is a divisional application with an application date of September 25, 2018, an application number of 201811115054.7, and the title of the invention "Kit for the detection of procalcitonin by spatial proximity chemiluminescence and its detection method". technical field [0002] The invention relates to the field of biotechnology, in particular to a kit for detecting procalcitonin by a spatial proximity chemiluminescence method, a detection method and application thereof. Background technique [0003] Procalcitonin (PCT) is composed of amino acids and belongs to the calcitonin propeptide substance without hormone activity, with good stability and a half-life of 25-30h. PCT is generally produced in thyroxine cells and forms calcitonin by specific proteolysis. Under pathological conditions, PCT is stimulated and induced by inflammatory cytokines and bacterial toxins, and secreted and produced in extrathyroxine tissues and organs. Its levels are reported...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/535G01N21/76
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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