Extraction buffer solution, rabbit brain extraction solution, PT detection reagent and PT detection kit

A technology for detection kits and detection reagents, which is applied in the field of medical detection and can solve problems such as easy precipitation and poor stability of PT reagents

Pending Publication Date: 2020-03-17
BEIJING SICCEEDER TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Excessively high solute concentration and the presence of a large amount of phospholipids make this PT reagent poor in stability, that is, it is easy to form precipitates

Method used

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  • Extraction buffer solution, rabbit brain extraction solution, PT detection reagent and PT detection kit
  • Extraction buffer solution, rabbit brain extraction solution, PT detection reagent and PT detection kit
  • Extraction buffer solution, rabbit brain extraction solution, PT detection reagent and PT detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] (1) Preparation of extraction buffer

[0039] 0.3% trisodium citrate, 0.3% anhydrous sodium acetate, adjust pH to 6.4 with acetic acid.

[0040] (2) Obtaining Rabbit Brain Extract

[0041] Dissolve 4.5% rabbit brain powder in the prepared extraction buffer (in g / mL, the mass volume ratio of rabbit brain powder to extraction buffer is 4.5:100), incubate at 39°C for 1 h, shake every 15 min Homogenize once, collect the supernatant after centrifugation at 3000RPM, which is the rabbit brain extract.

[0042] (3) Addition of lyoprotectant and heparin antagonist

[0043] Lyoprotectant, heparin antagonist and calcium ions were added to the rabbit brain extract. The protective agent is 0.5% bovine serum albumin, 1% glycine, the heparin antagonist is 0.03mg / ml polybrene, and the calcium ion is 15mM calcium chloride.

[0044] (4) freeze-drying, reconstitute with purified water after freeze-drying.

Embodiment 2

[0046] (1) Preparation of extraction buffer

[0047] 0.1% trisodium citrate, 1% anhydrous sodium acetate, adjust pH to 6.0 with acetic acid.

[0048] (2) Obtaining Rabbit Brain Extract

[0049] Dissolve 3% rabbit brain powder with the prepared extraction buffer (in g / mL, the mass volume ratio of rabbit brain powder to extraction buffer is 3:100), incubate at 33°C for 1.5h, every 15min Shake once, centrifuge at 3000RPM and collect the supernatant, which is the rabbit brain extract.

[0050] (3) Addition of lyoprotectant and heparin antagonist

[0051]Lyoprotectant, heparin antagonist and calcium ions were added to the rabbit brain extract. The protective agent is 5% peptone, 10% alanine, the heparin antagonist is 0.01 mg / ml protamine, and the calcium ion is 10 mM calcium lactate.

[0052] (4) freeze-drying, reconstitute with purified water after freeze-drying.

Embodiment 3

[0054] (1) Preparation of extraction buffer

[0055] 1% trisodium citrate, 0.1% anhydrous sodium acetate, adjust pH to 7.0 with acetic acid.

[0056] (2) Obtaining Rabbit Brain Extract

[0057] Dissolve 5% rabbit brain powder with the prepared extraction buffer (in g / mL, the mass volume ratio of rabbit brain powder to extraction buffer is 5:100), incubate at 45°C for 0.5h, every 15min Shake once, centrifuge at 3000RPM and collect the supernatant, which is the rabbit brain extract.

[0058] (3) Addition of lyoprotectant and heparin antagonist

[0059] Lyoprotectant, heparin antagonist and calcium ions were added to the rabbit brain extract. The protective agent is 3% bovine serum albumin, 5% glycine, the heparin antagonist is 0.1mg / ml polybrene, and the calcium ion is 25mM calcium carbonate.

[0060] (4) freeze-drying, reconstitute with purified water after freeze-drying.

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Abstract

The invention relates to the field of medical detection, in particular to an extraction buffer solution, a rabbit brain extraction solution, a PT detection reagent and a PT detection kit. The extraction buffer solution comprises citrate or phosphate, anhydrous sodium acetate and water. With adding of the electrolyte, the repulsive force of charges between phospholipids is increased, so that the PTreagent is not easy to precipitate. The stability is enhanced; and the repeatability during testing is improved.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to an extraction buffer, a rabbit brain extract, a PT detection reagent and a PT detection kit. Background technique [0002] Clinically, before the operation, in order to know whether the patient's hemostatic function is defective, to prepare in advance, to prevent hemorrhage during the operation and to be caught off guard, it is necessary to detect the patient's coagulation function, and the prothrombin time (PT) is one of the detection of coagulation function. one. PT is the main way to detect the lack of exogenous coagulation factors, it is to confirm the existence of congenital or acquired fibrinogen, prothrombin and coagulation factor V, VII, X defects or inhibitors, and it is also used to detect oral anticoagulation It is the preferred indicator for monitoring oral anticoagulants. [0003] The main component of PT reagent is tissue thromboplastin (i.e. a mixture of tissue f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/86
CPCG01N33/86
Inventor 丁重辉胡晓娟
Owner BEIJING SICCEEDER TECH CO LTD
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