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ck‑mb calibrator dilution and its application in clinical ck detection

A diluent and calibrator technology, applied in the CK-MB calibrator diluent and its application in clinical CK detection, can solve problems such as instability, improve accuracy, improve stability, and strengthen protein protection Effect

Active Publication Date: 2017-09-01
同昕生物技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] Since CK-MB exists in the form of a dimer and is extremely unstable, the calibrators of most company kits are stored in the form of freeze-dried products, which brings a lot of inconvenience in the process of clinical application.

Method used

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  • ck‑mb calibrator dilution and its application in clinical ck detection
  • ck‑mb calibrator dilution and its application in clinical ck detection
  • ck‑mb calibrator dilution and its application in clinical ck detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The pH value selection of embodiment 1.CK-MB calibrator dilution

[0049] 1. Preparation of CK-MB calibrator dilutions in different buffer systems

[0050] (1) 100ml CK-MB calibrator buffer solution (PBS)

[0051] Weigh sodium dihydrogen phosphate (NaH 2 PO 4 ) (Sinopharm, 20040799) 0.0624g, disodium hydrogen phosphate (Na 2 HPO 4 ) (Sinopharm, 100203008) 0.716g, sodium chloride (NaCl) (Sinopharm, 10019308) 0.8g, EDTA (Sigma, SE513402) 0.292g, reduced glutathione (Sigma, G4251) 0.0614g, fetal bovine serum ( Sigma, S12003C01) 10ml, zein (Sinopharm, XW901066) 5g. Add 60ml of ultrapure water, after completely dissolving, adjust the pH value to 7.4, and dilute to 100ml.

[0052] (2) 100ml CK-MB calibrator buffer (Tris)

[0053] Weigh Tris base (Amresco, ZA082601) 1.21g, EDTA (Sigma, SE513402) 0.292g, reduced glutathione (Sigma, G4251) 0.0614g, fetal bovine serum (Sigma, S12003C01) 10ml, zein (Sinopharm, XW901066) 5g. Add 60ml of ultrapure water, after completely di...

Embodiment 2

[0071] The protective agent selection of embodiment 2.CK-MB calibrator diluent

[0072] 1. Preparation of CK-MB calibrator dilutions containing different protective agents

[0073] (1) 100ml CK-MB calibrator buffer solution (the protective agent is zein)

[0074] Weigh sodium dihydrogen phosphate (NaH 2 PO 4 ) (Sinopharm, 20040799) 0.0624g, disodium hydrogen phosphate (Na 2 HPO 4 (Sinopharm, 100203008) 0.716g, sodium chloride (NaCl) (Sinopharm, 10019308) 0.8g, EDTA (Sigma, SE513402) 0.292g, reduced glutathione (Sigma, G4251) 0.0614g, fetal bovine serum (Sigma , S12003C01) 10ml, zein (Sinopharm, XW901066) 5g. Add 60ml of ultrapure water, after completely dissolving, adjust the pH value to 7.4, and dilute to 100ml.

[0075] (2) 100ml CK-MB calibrator buffer solution (protective agent is BSA)

[0076] Weigh sodium dihydrogen phosphate (NaH 2 PO 4 ) (Sinopharm, 20040799) 0.0624g, disodium hydrogen phosphate (Na 2 HPO 4(Sinopharm, 100203008) 0.716g, sodium chloride (NaCl...

Embodiment 3

[0093] The screening of the optimal concentration of each component in embodiment 3.CK-MB calibrator diluent

[0094] 1. Preparation of CK-MB calibrator dilutions containing different concentrations of EDTA

[0095] (1) 100ml CK-MB calibrator buffer (without EDTA)

[0096] Weigh sodium dihydrogen phosphate (NaH 2 PO 4 ) (Sinopharm, 20040799) 0.0624g, disodium hydrogen phosphate (Na 2 HPO 4 (Sinopharm, 100203008) 0.716g, sodium chloride (NaCl) (Sinopharm, 10019308) 0.8g, reduced glutathione (Sigma, G4251) 0.0614g, fetal bovine serum (Sigma, S12003C01) 10ml, zein (Sinopharm , XW901066) 5g. Add 60ml of ultrapure water, after completely dissolving, adjust the pH value to 7.4, and dilute to 100ml.

[0097] (2) 100ml CK-MB calibrator buffer (5mM EDTA)

[0098] Weigh sodium dihydrogen phosphate (NaH 2 PO 4 ) (Sinopharm, 20040799) 0.0624g, disodium hydrogen phosphate (Na 2 HPO 4 (Sinopharm, 100203008) 0.716g, sodium chloride (NaCl) (Sinopharm, 10019308) 0.8g, EDTA (Sigma, S...

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Abstract

The invention relates to a medical science detection reagent, and especially relates to a CK-MB calibrating substance dilution solution and an application thereof in clinic CK detection. The CK-MB calibrating substance dilution solution is prepared through adding corn proteins with the final concentration of 0.01-0.12g / ml, glutathione with the final concentration of 0.5-4mM, fetal calf serum with the final concentration of 3-20% (V / V) and EDTA with the final concentration of 3-20mM to a buffering solution. The dilution solution maximally improves the stability of a CK-MB calibrating substance, solves the unstable problem of present CK-MB calibrating substances, improves the clinic CK detection accuracy, and provides accurate basis for diagnosis of clinic related diseases.

Description

technical field [0001] The invention relates to a medical detection reagent, especially a CK-MB calibrator diluent and its application in clinical CK detection. [0002] technical background [0003] Creatine kinase (CK) exists in the form of dimers and plays an important role in the generation and application of high-energy phosphate bonds in contractile tissues. Moreover, in the cytoplasm, the ATP produced by mitochondria is delivered to the site of use through creatine phosphocreatine, and creatine kinase plays the role of "shuttle transport". The "shuttle hypothesis" has been confirmed in some tissues. In the distal convoluted tubule of the kidney, although it does not contract, it requires a large amount of ATP to supply the sodium and potassium pumps to maintain the ion gradient. The chemical reaction catalyzed by CK is: creatine (Creatine) + ATP generates phosphocreatine (creatine phosphate, CP) and ADP under the catalysis of CK. The ATP produced by the mitochondria ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/96G01N33/573G01N33/531
CPCG01N33/531G01N33/573
Inventor 肖智李全焦守恕
Owner 同昕生物技术(北京)有限公司