What are the characteristics of the antithrombin III assay kit

The core characteristics of the antithrombin III assay kit are reflected in the scientific nature of the detection principle, the accuracy of the reagent composition, the superiority of performance indicators, the convenience of the operation process, and the wide range of application scenarios. Specifically, the following:

1. Detection principle: Chromogenic substrate method, scientifically reliable

The antithrombin III assay kit uses the chromogenic substrate method for detection. Add a quantitative amount of thrombin to the sample, and antithrombin III (AT-III) forms a 1:1 complex with thrombin to inactivate the corresponding thrombin. The remaining thrombin acts on the chromogenic substrate, releasing chromogenic groups, and the increase in absorbance is measured at a wavelength of 405nm. The change in absorbance is negatively correlated with AT-III activity, and the activity of AT-III can be calculated based on the standard curve. This principle is scientifically reliable and provides a solid foundation for the accuracy of detection results.

2. Reagent composition: precise ratio to ensure effectiveness

The components of the reagent kit have been carefully screened and optimized, including thrombin reagent, thrombin substrate, and dilution buffer.

. Taking a certain brand of reagent kit as an example, its thrombin reagent is 25nCat freeze-dried bovine thrombin, the thrombin substrate is H-D-HHT-L-Ala-L-Arg-pNA · 2AcOH, and the dilution buffer contains Tris HCl, NaCl, NaEDTA, heparin sodium and other components. The precise ratio of these ingredients ensures the stability and accuracy of the reagent kit during the detection process.

III. Performance indicators: superior and outstanding, meeting requirements

Accuracy: The accuracy of the reagent kit is good, with high sensitivity and few interference factors.

. For example, a certain brand of reagent kit has high precision, CV (coefficient of variation)<10%, and requires a small sample size, only a few microliters of sample are needed to complete the detection. Specificity: The kit has high specificity and does not cross react with other soluble structural analogues. This ensures the accuracy and reliability of the test results, avoiding the occurrence of false positives or false negatives.

Repeatability: The reagent kit has good repeatability, with intra plate and inter plate coefficients of variation both less than 15%.

. This means that the results of testing conducted at different times and batches can be consistent, providing reliable basis for clinical diagnosis and treatment. Stability: The reagent kit exhibits good stability during storage and use. For example, the R1 and R2 reagents of a certain brand of reagent kit can be stored for 8 days at+2~+8 ℃ after opening, while the AT-III reagent kit can be stored for 18 months at+2~+8 ℃. This extends the service life of the reagent kit and reduces the cost of use.

Fourthly, the operation process is simple, fast, and easy to master.

The operation process of the reagent kit is simple, fast, and easy to master.

. Generally speaking, the operational steps include sample processing, sample addition, incubation, washing, color development, and reaction termination. Taking the ELISA detection kit as an example, the operating steps are as follows:

Sample processing: Collect samples such as serum and plasma, and perform appropriate processing (such as centrifugation of serum).

Sample addition: Add the sample, standard, and HRP labeled detection antibody in sequence to the pre coated micro well with antithrombin III (AT-III) antibody.

. Incubation: Cover the plate with a sealing film and incubate at 37 ℃ for a certain period of time (such as 30 minutes).

Washing: Carefully remove the sealing film, discard the liquid, shake dry, fill each hole with washing solution, let it stand for a certain period of time, discard, repeat this multiple times, pat dry.

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Color development: Add color developer A to each well first, then add color developer B, gently shake and mix, and develop color at 37 ℃ in the dark for a certain period of time (such as 15 minutes).

. Termination reaction: Add termination solution to each well to terminate the reaction (at this point, the blue color turns yellow).

Measurement: Measure the absorbance (OD value) of each well in sequence using blank air conditioning zero and 450nm wavelength, and calculate the sample concentration.

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Fifth, application scenarios: Widely diverse, meeting clinical needs

The antithrombin III assay kit has a wide range of clinical applications.

. It can be used for the diagnosis and monitoring of genetic antithrombin deficiency, pre thrombotic state and thrombotic diseases, synthesis reduction (such as severe liver disease), and other diseases. For example:

Hereditary antithrombin deficiency: caused by AT gene mutations, showing an autosomal dominant inheritance pattern, mostly accompanied by a family history. Surgery, trauma, and infection are the main causes of thrombosis in congenital AT-III deficiency. By detecting AT-III activity, it can assist in the diagnosis of this disease. Pre thrombotic state and thrombotic diseases: such as DIC hypercoagulability, myocardial infarction, angina pectoris, cerebrovascular disease, pregnancy, deep vein thrombosis, nephrotic syndrome, tumors, etc. In these disease states, blood anticoagulation is weakened and AT-III activity is reduced. By detecting AT-III activity, the coagulation status of patients can be evaluated, providing a basis for treatment.

Reduced synthesis: such as severe liver disease.

. The liver is the main site for the synthesis of AT-III, and in severe liver disease, the synthesis and activity of AT-III decrease. By detecting AT-III activity, it can assist in the diagnosis of liver disease and assess its severity.