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A rapid antithrombin III detecting kit based on interaction between thrombin and a chromogenic substrate and a detecting method thereof

A detection kit and antithrombin technology, which is applied in the direction of material analysis through observation of the influence on chemical indicators, and analysis through chemical reactions of materials, etc., which can solve problems such as inability to carry out detection items and lack of detection methods , to achieve the effect of improving naked eye visibility and simple operation

Inactive Publication Date: 2016-04-06
鲁翌
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The current diagnostic technology is developing in two directions of large-scale integration and simple and rapid development. The chromogenic substrate method is widely used in the in vitro diagnostic industry, especially in the field of thrombosis and hemostasis, and the accuracy of the detection results is good. However, the existing The test of chromogenic substrate method must use matching instruments to realize the detection
However, in many cases, if the conditions are not allowed (when there is no instrument or the instrument does not have this function), it is impossible to carry out the detection project in this area
When it is only necessary to simply and quickly judge the approximate content or only need to qualitatively judge the positive and negative, the existing detection method of chromogenic substrate method is relatively deficient.

Method used

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  • A rapid antithrombin III detecting kit based on interaction between thrombin and a chromogenic substrate and a detecting method thereof
  • A rapid antithrombin III detecting kit based on interaction between thrombin and a chromogenic substrate and a detecting method thereof
  • A rapid antithrombin III detecting kit based on interaction between thrombin and a chromogenic substrate and a detecting method thereof

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Effect test

Embodiment 1

[0030] Embodiment 1 detection kit and detection method thereof

[0031] An antithrombin III rapid detection kit based on the interaction between thrombin and a chromogenic substrate, comprising R1 reagent, R2 reagent, stop solution and chromogenic assay card.

[0032] The R1 reagent is: thrombin 5000IU / L, heparin 3000IU / L, Tris-glycine 50mM, pH8.4, sodium chloride 175mM, mannitol 5wt% (mass concentration), Na 2 EDTA0.2wt%, BSA2wt% and sodium azide.0.1wt%. Freeze-dried in aliquots and reconstituted with an equal volume of purified water before use.

[0033] The R2 reagent is: chromogenic substrate Su60.25mg / ml, Tris-HCl50mM, pH8.4, Tween-201wt%, PEG80001wt%, trehalose3.5wt%, sodium azide0.01wt%.

[0034] The stop solution is: 50 wt% acetic acid, 0.01 wt% sodium azide.

[0035] Described chromogenic assay card (concrete structure such as figure 1 ) is made of white PVC plastic, the volume of the color measuring card: 7cm in length, 2cm in width, and 0.5cm in thickness; there...

Embodiment 2

[0043] Embodiment 2 kit detection accuracy

[0044] The reference plasma is diluted into a series of concentrations (100%, 75%, 50%, 25%, 0%) with physiological saline, and then the antithrombin III activity is detected by the kit of the present invention, while using an existing blood coagulation Analyzer detection, comparative detection results are shown in Table 1:

[0045] The AT-III activity is detected by different methods after the reference plasma is diluted in proportion in Table 1:

[0046]

[0047] Collect the patient's plasma as a test sample, and use the kit-based detection method of the present invention to detect, and simultaneously detect with an existing coagulation analyzer, and the comparative detection results are as follows in Table 2:

[0048] Table 2 Patient's plasma is tested for AT-III activity by different detection methods:

[0049]

[0050]

[0051] The test results in Table 1 and Table 2 show that the antithrombin III activity can be det...

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Abstract

The invention belongs to the field of biological medicine in-vitro diagnosis, and particularly relates to a rapid antithrombin III detecting kit based on interaction between thrombin and a chromogenic substrate and a detecting method thereof. The kit comprises R1 reagents, an R2 reagent, a stop buffer and a chromogenic measuring card. The R2 reagent comprises a chromogenic substrate reagent. The chromogenic measuring card is provided with a colorimetric reference belt and a reaction cell. The colorimetric reference belt is provided with six color zones which have colors from deep to light and are respectively corresponding to 20%, 40%, 60%, 80%, 100% and higher than 100% of activity of the antithrombin III. Naked-eye distinctiveness of a chromogenic reaction is improved by improving a formula of the chromogenic substrate and the activity of the antithrombin III and colors distinguishable with naked eyes are correlated to obtain the colorimetric reference belt distinguishable by human eyes, and therefore the activity of the antithrombin III can be rapidly and semiquantitatively detected only by naked-eye determination without the need of using matched instruments.

Description

technical field [0001] The invention belongs to the field of biomedical in vitro diagnostic reagents, in particular to an antithrombin III rapid detection kit and a detection method based on the interaction between thrombin and a chromogenic matrix. Background technique [0002] The detection principle of the chromogenic substrate method is as follows: artificially synthesize a compound that can be catalyzed and cleaved by the thromboplastin to be tested, and this compound is connected to a chromogenic substrate. There is a color change in the test sample, and the activity of the tested thromboplastin can be deduced according to the color change. Chromogenic substances are generally selected to connect p-nitroaniline (pNA). [0003] At present, this method is commonly used in biochemical analyzers, spectrophotometers and other equipment. The free pNA is yellow, and its measurement wavelength is generally 405nm. At this wavelength, other substances absorb less than 1% of th...

Claims

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Application Information

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IPC IPC(8): G01N21/78
CPCG01N21/78
Inventor 鲁翌田树伟吕学敏董小蓉张隽
Owner 鲁翌
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