The testing method of adenosine deaminase

A technology for detecting reagents and purine nucleoside phosphorylase, which is applied in the field of medical testing, can solve problems such as large differences in test results, unfavorable clinical test results, and troubles in clinical diagnosis and treatment, and achieve stable detection methods, overcome non-specific reactions, and clinically The effect of medical convenience

Inactive Publication Date: 2017-01-04
GUANGDONG HOSPITAL OF TRADITIONAL CHINESE MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the current clinical ADA detection methods all adopt the detection principle of four-step enzyme coupling, the performance is uneven, and there are differences in the reagent formula and detection process of different manufacturers, the detection results of different systems vary greatly, and there is no uniform The reference interval is not conducive to the consistency of clinical test results, which brings many troubles to clinical diagnosis and treatment

Method used

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  • The testing method of adenosine deaminase
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  • The testing method of adenosine deaminase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] 1. Reagent composition and concentration

[0067] Reagent 1: Tris-HCl (80.0mmol / L, pH6.6), disodium hydrogen phosphate (Na 2 HPO 4 , 10.3mmol / L), 4-aminoantipyrine (4-AA, 1.8mmol / L), purine nucleoside phosphorylase (PNP, 37°C 2760U / L), xanthine oxidase (XOD, 37°C 600U / L), peroxidase (POD, 590U / L at 37°C), ascorbate (2306U / L at 37°C), NaN 3 (0.2mmol / L), TritonX-100 (0.76% of the total volume of reagent 1), EDTA (0.2mmol / L);

[0068] Reagent 2 (starting reagent): Tris-HCl (80.0 mmol / L, pH 6.6), adenosine (12.2 mmol / L), EHSPT (2.0 mmol / L).

[0069] 2. Detection conditions

[0070]Detection temperature: 37±0.1°C; detection wavelength: 550±1nm; wave width: 10mm±0.01mm, optical path ≤ 2nm, detection points ≥ 6; incubation time: 300s; delay time: 120s; monitoring time: 180s.

[0071] 3. Test sample: serum.

[0072] 4. A detection method for adenosine deaminase, comprising the following steps:

[0073] 1) Add 180ul reagent 1 and 5ul serum samples into the cuvette, mix we...

Embodiment 2

[0079] The performance parameter that embodiment 2 adenosine deaminase detects

[0080] The performance parameters of the present technology were determined according to the method of Example 1.

[0081] The result is as follows:

[0082] Intra-assay precision: ≤1%, period precision: ≤2%;

[0083] Linear range: 0.3-286U / L;

[0084] Sensitivity: The activity of the detection standard substance exceeds the given target value by about 30%.

[0085] Anti-interference ability: triglyceride ≤ 3.39mmol / L (about 20 times the normal adult level), hemoglobin ≤ 3.75g / L (visible hemolysis is 300mg / dl), bilirubin < 85.5umol / L, vitamin C ≤ 40mg / L (the content of vitamin C in normal human serum is ng / dl level), glycyrrhetinic acid and deoxycholic acid ≤ 20mg / L, baicalein ≤ 37.5mg / L will not interfere with the method.

[0086] The linear range diagram of the detection method is shown in the appendix figure 2 . figure 2 The linear coefficient of the curve is significant, and the correl...

Embodiment 3

[0088] Embodiment 3 repeatability test

[0089] Next, the method established by the present invention is tested for repeatability, and the intra-batch precision and period precision are calculated respectively.

[0090] Serum samples (high and low concentrations) with no visible lipemia, hemolysis, jaundice and negative infectious indicators were selected. Intra-batch precision: each sample is tested 10 times in a batch, and the mean, standard deviation and coefficient of variation (CV) are calculated. The precision during the period is: each sample is tested 3 times a day, and the mean and standard deviation are calculated for 12 consecutive days. and coefficient of variation (CV).

[0091] The results were intra-assay precision: ≤ 1%, inter-assay precision: ≤ 2%.

[0092] The intra-assay and inter-interval repeatability of the method are good.

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Abstract

The invention discloses the testing method of adenosine deaminase. The testing reagent of adenosine deaminase is made up of the following two parts: Reagent 1 includes disodium hydrogen phosphate 8.5-12 mmol / L, aminoantipyrine 1.5-2.2 mmol / L, purine nucleoside phosphorylase 2500-2900 U / L, xanthine oxidase 500-700 U / L, peroxidase 510-710 U / L, ascorbinase 1800-2800 U / L, certain stability reagent and certain assistant factors. Reagent 2 includes adenosine 10-14 mmol / L and EHSPT 1.5-2.5 mmol / L. The testing reagent formula of adenosine dehydrogenase in this invention and the confirmation of the testing parameters and referential range are good for the consistency of the clinical testing results. it will bring a lot of benefits to the clinical diagnosis. The sensitivity of this method is very high and it is superior to the similar products sold on the market.

Description

technical field [0001] The invention belongs to the field of medical testing, and in particular relates to a method for detecting adenosine deaminase. Background technique [0002] Adenosine deaminase (ADA) is an essential nucleic acid catabolic enzyme in the body and has important value in clinical laboratory diagnosis. At present, enzyme detection mostly adopts the continuous monitoring method of enzyme coupling principle to determine the concentration of enzyme catalytic activity in the sample. This method has the advantages of simple, fast, non-toxic and non-radioactive, and is widely used in clinical practice. Although the current clinical ADA detection methods all adopt the detection principle of four-step enzyme coupling, the performance is uneven, and there are differences in the reagent formula and detection process of different manufacturers, the detection results of different systems vary greatly, and there is no unified standard. The reference interval is not co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/34C12Q1/48C12Q1/28C12Q1/26
CPCC12Q1/34C12Q1/26C12Q1/28C12Q1/48
Inventor 黄宪章韩丽乔庄俊华王建兵林海标张乔轩
Owner GUANGDONG HOSPITAL OF TRADITIONAL CHINESE MEDICINE
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