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Kit for detecting free semen elastinase and application thereof

An elastase and detection kit technology, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of unsuitable popularization and application, complicated operation steps, and many sample adding procedures, and achieves short detection time, Accurate and repeatable test results

Pending Publication Date: 2018-01-12
南京欣迪生物药业工程有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can only detect the level of NE inhibited by α1-antitrypsin, but the level of free NE that really plays a role in the inflammatory process cannot be reflected. Therefore, this method has obvious limitations in reflecting the degree of inflammation in the male reproductive system. At the same time, this method is a manual method, with many sample addition procedures and complicated operation steps, which is not only time-consuming and labor-intensive, but also has relatively large human errors, so this method is not suitable for clinical application.

Method used

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  • Kit for detecting free semen elastinase and application thereof
  • Kit for detecting free semen elastinase and application thereof
  • Kit for detecting free semen elastinase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Semen free elastase detection kit, including the following components:

[0039] R1 reagent: dimethyl sulfoxide dissolved in Hepes buffer solution, 0.75g polyoxyethylene lauryl ether, 0.3mol sodium chloride and 0.03mol N-(2-hydroxyethyl)piperazine-N' per liter of Hepes buffer solution - 2-ethanesulfonic acid, adjust its pH to 8.0 to obtain a Hepes buffer solution, add 19 ml of dimethyl sulfoxide to this buffer solution (1 L) to obtain R1 reagent.

[0040] R2 reagent: After completely dissolving N-methoxysuccinyl-propionamide-propionylamide-prolylamide-valine-4-nitroanilide in dimethyl sulfoxide, add Hepes buffer solution ( Same as in R1) dissolved to obtain R2 reagent, containing 3.7mmol N-methoxysuccinyl-propionylamide-propionylamide-prolineamide-valine-4-nitroanilide in every liter of R2 reagent, 19ml of dimethylsulfoxide and 0.0245mol of N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid.

[0041] The calibrator is an elastase solution, wherein the elastase activ...

Embodiment 2

[0045] Semen free elastase detection kit, including the following components:

[0046] R1 reagent: dimethyl sulfoxide dissolved in Hepes buffer solution, containing 1.2g polyoxyethylene lauryl ether, 0.1mol sodium chloride and 0.05mol N-(2-hydroxyethyl)piperazine-N per liter of Hepes buffer solution '-2-ethanesulfonic acid, adjust its pH=9.0 to obtain Hepes buffer solution, add 30ml dimethyl sulfoxide to this buffer solution (1L) to obtain R1 reagent.

[0047] R2 reagent: After completely dissolving N-methoxysuccinyl-propionamide-propionylamide-prolylamide-valine-4-nitroanilide in dimethyl sulfoxide, add Hepes buffer solution ( Same as in R1) dissolved to obtain R2 reagent, containing 5.0mmol N-methoxysuccinyl-propionylamide-propionylamide-proylamide-valine-4-nitroanilide in every liter of R2 reagent, 30ml dimethyl sulfoxide, 0.0485mol N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid.

[0048] The calibrator is an elastase solution, wherein the elastase activity is 10 U...

Embodiment 3

[0052] Semen free elastase detection kit, including the following components:

[0053] R1 reagent: dimethyl sulfoxide dissolved in Hepes buffer solution, containing 0.3g polyoxyethylene lauryl ether, 0.5mol sodium chloride and 0.01mol N-(2-hydroxyethyl)piperazine-N per liter of Hepes buffer solution '-2-ethanesulfonic acid, adjust its pH=7.0 to obtain Hepes buffer solution, add 8ml dimethyl sulfoxide to this buffer solution (1L) to obtain R1 reagent.

[0054] R2 reagent: After completely dissolving N-methoxysuccinyl-propionamide-propionylamide-prolylamide-valine-4-nitroanilide in dimethyl sulfoxide, add Hepes buffer solution ( Same as in R1) dissolving to obtain R2 reagent, containing 2.4mmol N-methoxysuccinyl-propionylamide-propionylamide-prolineamide-valine-4-nitroanilide in every liter of R2 reagent, 8ml dimethyl sulfoxide, 0.0097mol N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid.

[0055] The calibrator is elastase solution, wherein the activity of elastase is 5U / ...

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Abstract

The invention discloses a kit for detecting free semen elastinase and an application thereof. The kit comprises a reagent R1, a reagent R2, a calibrating material, a quality control material and a sample treating solution, wherein the R1 reagent is a Hepes buffering solution containing dimethyl sulfoxide; the R2 reagent is the Hepes buffering solution containing N-methoxy succinyl-propanamide-propanamide-prolyl-valine-4-propionamide and dimethyl sulfoxide; the calibrating material is an elastinase solution; the quality control material is the freeze-dried powder containing elastinase; the sample treating solution is a triton X-100 solution. The kit disclosed by the invention is used together with a full-automatic biochemical analyzer for detecting the free semen elastinase; when the free semen elastinase is detected, the reaction time is short, the required reagent dosage and sample volume are less, the linear scope is wide and the result accuracy and precision are high; the kit disclosed by the invention is suitable for clinically detecting the samples in large scale; the whole reaction process is monitored through the change of the reaction curve.

Description

technical field [0001] The invention relates to the second type of in vitro diagnostic reagents, in particular to a semen free elastase detection kit and application thereof. Background technique [0002] Elastase is an enzyme capable of hydrolyzing elastin in the body, and the elastase present in neutrophils is called neutrophil elastase (NE). NE is an important neutral proteolytic enzyme, which directly participates in various physiological and pathological processes in the body, and plays an important mediator role in many aspects such as infectious diseases, tissue damage and inflammation. The level of elastase in various body fluids or tissues can well reflect the severity of tissue organ damage and is a sensitive indicator for judging its prognosis. Under normal physiological conditions, the amount of elastase secreted by macrophages and neutrophils is very small, and it constantly interacts with anti-protease systems such as α1-antitrypsin (α1-AT) and α2-macroglobuli...

Claims

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Application Information

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IPC IPC(8): C12Q1/37
Inventor 陆金春宰世凤刘园园陈晓慧
Owner 南京欣迪生物药业工程有限责任公司
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