Immunofluorescent quantitative test strip detecting creatine kinase isoenzyme mass

A creatine kinase and immunofluorescence technology, applied in the field of immunological detection, can solve the problems of complicated operation, poor reagent stability, poor detection accuracy, etc., and achieve the effects of high chromatographic recovery rate, high detection accuracy and high correlation coefficient.

Inactive Publication Date: 2018-05-11
ZHONGSHAN CHUANGYI BIOCHEM ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Enzyme-linked immunosorbent assay detection is time-consuming and complicated to operate, and is not suitable for rapid detection in large quantities
After the latex-enhanced immunoturbidimetric reaction, latex particles will precipitate, which is not conducive to the cleaning of the instrument, and th

Method used

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  • Immunofluorescent quantitative test strip detecting creatine kinase isoenzyme mass
  • Immunofluorescent quantitative test strip detecting creatine kinase isoenzyme mass
  • Immunofluorescent quantitative test strip detecting creatine kinase isoenzyme mass

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A preferred formula of the present invention: the mass concentration of PB is 20mM, the percentage concentration of BSA is 1.8%, the percentage concentration of Tween-80 is 0.5%, the percentage concentration of glucose is 0.5%, and the percentage concentration of glycine is 2 %, the percentage concentration of PEG4000 is 1%, the percentage concentration of PEG20000 is 1.5%, the percentage concentration of Proclin300 or sodium azide is 0.03%, pH7.5.

[0026] Preparation method of storage solution: Weigh 0.25g of glucose and dissolve it in 45mL of pure water. Add 1mL of 100mM PB stock solution prepared in advance, shake and mix well. Add 1g of glycine, 0.5g of PEG4000, 0.75g of PEG20000 and 0.9g of BSA successively, shake and mix well; add 0.25mL Tween-80 with a sample gun, mix well repeatedly; add 15μL of Proclin300, shake and mix well; adjust the pH to 7.5 with 1M HCl . Finally, the volume was adjusted to 50 mL at room temperature and sterilized by filtration to obtai...

Embodiment 2

[0029] The preparation process of creatine kinase isoenzyme quality immunofluorescence quantitative test strip is as follows:

[0030] (1) Preparation of storage solution: the mass concentration of PB is 20mM, the percentage concentration of BSA is 1.8%, the percentage concentration of Tween-80 is 0.5%, the percentage concentration of glucose is 0.5%, and the percentage concentration of glycine is 2%, the percentage concentration of PEG4000 is 1%, the percentage concentration of PEG20000 is 1.5%, the percentage concentration of Proclin300 is 0.03%, and the pH7.5 is prepared according to the above formula, mixed and then filtered and sterilized to prepare the storage solution;

[0031] (2) Preparation of the sample pad: Soak the glass fiber membrane with the sample pad treatment solution for 10 minutes, place in a drying room at 37°C and 30% humidity, and dry for 3 hours to prepare the sample pad for use;

[0032](3) Preparation of the bonding pad: Soak the glass cellulose memb...

Embodiment 3

[0046] Prepare the control storage solution, the specific formula is: the mass concentration of PB is 50mM, the percentage concentration of BSA is 1%, the percentage concentration of Tween-80 is 1%, the percentage concentration of glucose is 1%, the percentage concentration of glycine 0.5%, the percentage concentration of PEG4000 is 2%, and the percentage concentration of Proclin300 is 0.3%.

[0047] The preferred storage solution prepared in Example 1 (hereinafter referred to as storage solution 2) is compared with the reference storage solution (hereinafter referred to as storage solution 1) for stabilizing performance and precision, and the implementation method and results are as follows:

[0048] Use storage solution 1 and storage solution 2 to spray and dry the fluorescent microspheres labeled with creatine kinase isoenzyme quality (CK-MB) monoclonal antibody by the same process, assemble them into 40 test strips, put them in aluminum foil bags, and dry them. agent seal....

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Abstract

The invention discloses an immunofluorescent quantitative test strip detecting the creatine kinase isoenzyme mass. The prepared creatine kinase isoenzyme immunofluorescent quantitative test strip is high in standard curve correlation coefficient, high in detection accuracy, good in stability, high in degree of precision and high in sensitivity. Accordingly, an applied stock solution has the high chromatographic recovery rate, almost no fluorescent microsphere remains on a conjugate pad or runs to absorbent paper on the test strip, and almost all the fluorescent microspheres are captured. The test strip is suitable for clinical detection, and the test strip has the advantages of being convenient to operate, rapid in reaction, high in sensitivity, high in specificity, suitable for field detection, economical, practical and the like.

Description

technical field [0001] The invention relates to the field of immunological detection, and more specifically relates to an immunofluorescence quantitative test strip for detecting the quality of creatine kinase isoenzymes. Background technique [0002] Creatine Kinase (Creatine Kinase, CK) (ATP: Creatine N-phosphotransferase EC2.7.3.2) usually exists in the cytoplasm and mitochondria of the heart, muscle and brain of animals, and is an enzyme related to intracellular energy operation and muscle contraction. , an important kinase directly related to adenosine triphosphate (ATP) regeneration1,2, which reversibly catalyzes the transphosphorylation reaction between creatine and ATP. Creatine kinase has four isozyme forms: muscle (MM), brain (BB), hybrid (MB) and mitochondrial (MiMi). The increase of creatine kinase isoenzyme in the human body has the following conditions: acute myocardial infarction (CK-MB>0.03, up to 0.12-0.38), hypothyroidism, cerebrovascular disease, pulmo...

Claims

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Application Information

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IPC IPC(8): G01N33/533
CPCG01N33/533
Inventor 陈润文何平李冰肖丝尹周琼华
Owner ZHONGSHAN CHUANGYI BIOCHEM ENG
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