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Application of kit for detecting serum amyloid protein A by spatial proximity chemiluminescence method and detection method

A technology of chemiluminescence method and detection kit, which is applied in the direction of chemiluminescence/bioluminescence, analysis through chemical reaction of materials, and measurement devices, which can solve the problems of unstable detection results, many influencing factors, and poor repeatability, etc. problems, to achieve the effect of eliminating luminous interference, simple production process, and easy full automation

Pending Publication Date: 2021-10-19
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

Its operation is complicated and there are many influencing factors, resulting in unstable test results and poor repeatability

Method used

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Examples

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Effect test

preparation example Construction

[0031] First, the preparation of calibration

[0032] (1) Preparation of the calibration dilution: Tie a dihydrogen phosphate 14.1g, sodium phosphate (2H) 2O) 3.0g, additional water dissolution, proClin-3000.5 ~ 1 mL, mix, add ultra-pure water to 1000mL to obtain a calibration dilution, 2 to 8 ° C storage spare.

[0033] (2) Preparation of calibration: The concentration of the calibration is 0, 15 ng / ml, 45 ng / ml, 135 ng / ml, 400 ng / ml, 800 ng / ml, and the purified serum amylin A is diluted to the corresponding concentration with calibration dilution. Survey standby 2 ~ 8 ° C.

[0034] Second, the preparation of enzyme labeled

[0035] (1) It is weighing 5 mg of HRP in 1 ml of distilled water, and 0.2 ml of a new 0.1 m perionic acid solution solution was added to the superhecent, stirring was stirred at room temperature for 20 min, and the above solution was loaded into the dialysis bag, and 1 mMPH4. 4 sodium acetate buffer dialysis, 4 ° C overnight.

[0036] (2) Add 20 μl...

Embodiment 1

[0053] This embodiment provides a detection kit for serum amyloid A, comprising: an enzyme standard, a light emitting label, auxiliary agent, a trigger, and a calibrator; wherein the calibration comprises no less than 6 different concentrations of SAA antigen. Calibration and 0.1M phosphate buffer; the enzyme-based instrument includes a peroxidase-labeled SAA detection antibody and 0.05 M phosphate buffer; the light emitting label includes 9,10-dihydroacid-labeled SAA capture antibody and 0.05 M Tris buffer; components of the auxiliary agent include luminescent adjuvants and pH 6.0 citrate buffer; trigger agents use 0.05 mph 8.0 Tris-HCl buffer.

[0054] First, the preparation of calibration

[0055] (1) Preparation of the calibration dilution: Tie a dihydrogen phosphate 14.1g, sodium phosphate (2H) 2 O) 3.0g, additional water dissolved, proClin-3000.8mL, after mixing, add ultra-pure water to 1000mL to obtain a calibration dilution, 4 ° C storage spare.

[0056] (2) Preparation of...

Embodiment 2

[0070] This embodiment provides a method of detecting serum amyloid A using serum amyloid A detection kits to detect serum ammaid proteins a, including steps:

[0071] S1: 25 μl of standard, 25 μl of peroxidase-labeled SAA detection antibody and 25 μL of luminescent label are added to a chemiluminescence reaction tube.

[0072] S2: The thermostat reaction was 15 min at 37 ° C.

[0073] S3: 5 μL of adjuvant is added to the chemiluminescence reaction tube, and the oscillation is mixed, and the trigger is added to 75 μl, and the signal value is detected immediately after the trigger is added.

[0074] S4: The concentration and light emitting value of the calibration are fitted to calculate the sample concentration through the sample light value.

[0075] The present invention adopts a space adjacent chemiluminescence method to detect serum amyloid A, and the sensitivity can reach 1 mg / L. The results of SAA testing have high clinical significance for viral infection, showing: 99% ec...

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Abstract

The invention relates to a kit for detecting serum amyloid protein A by a space proximity chemiluminescence method and a detection method thereof. The kit comprises an enzyme marker, a luminous marker, an auxiliary agent, a triggering agent and a calibrator, wherein the calibrator comprises calibrators of SAA antigens with different concentrations and 0.1 M phosphate diluent; the enzyme marker comprises a peroxidase-labeled SAA detection antibody and a 0.05 M phosphate buffer solution; the luminous marker comprises a 9, 10-dihydroacridine labeled SAA capture antibody and a 0.05 M Tris buffer solution; and the auxiliary agent comprises a luminous auxiliary agent and a citrate buffer solution. According to the invention, a reagent space proximity luminescence analysis and detection technology is adopted as a real homogeneous chemiluminescence technology, and has the advantages of no need of cleaning, no need of a carrier, no coating process, high detection sensitivity and strong specificity, so that the detection result is more real and credible; and meanwhile, the reaction time and the reaction steps are optimized, so that the operation is simpler.

Description

[0001] The present application is the application date for September 25, 2018, and the application number is 20181115981.9, and the invention is a "Space adjacent chemiluminescence method to detect serum ammaid kit and detecting method". Technical field [0002] The present invention relates to the field of biotechnology, and more particularly to a kit of a space adjacent chemiluminescence detection serum ammaid protein A and its detection method thereof. Background technique [0003] Infectious diseases are clinically common diseases and multi-hazardous diseases, mainly inflammatory responses caused by pathogens such as bacteria, viruses and / or fungi, mostly with fever symptoms. The anti-bacterial drug is cured and saved many patients, and its unreasonable application and abuse also led to many bad consequences, such as bacteria, severe drug resistance. [0004] Serum amyloid is a polypeptide consisting of 104 amino acids. Its relative molecular mass at the time of natural stat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N33/577G01N33/535
CPCG01N21/76G01N33/535G01N33/577
Inventor 奚伟红史兵伟廖鸳鸯朱丹丹
Owner 无锡壹闪生物科技有限公司
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