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Test strip, preparation method thereof and application of test strip to combined detection of microalbuminuria and beta2 microglobulin

A technology of β2 microglobulin and albumin, which is applied in the field of biochemical detection, can solve the problems of the influence of detection results, low detection limit, and low detection limit, and achieve the effect of being suitable for on-site detection, improving accuracy, and strong specificity

Active Publication Date: 2017-02-22
SINOCARE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the low detection limit of the double-antibody sandwich method has brought troubles to the detection of MAU and β2-MG
The detection limit is too low, so that the urine sample needs to be diluted with a specific diluent before using the test strip, otherwise the test result of healthy people will also be positive
However, the dilution must be carried out according to a specific multiple, which not only increases the complexity of the operation, but also easily affects the detection results due to operational errors during the dilution process.

Method used

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  • Test strip, preparation method thereof and application of test strip to combined detection of microalbuminuria and beta2 microglobulin
  • Test strip, preparation method thereof and application of test strip to combined detection of microalbuminuria and beta2 microglobulin
  • Test strip, preparation method thereof and application of test strip to combined detection of microalbuminuria and beta2 microglobulin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Preparation of colloidal gold particles: Take 1L of water and put it into a round-bottomed flask, place it in an electric heating mantle and heat it to boiling, take 15mL-20mL of 1% trisodium citrate and add it to boil for 2 minutes, add 10mL of 2%-4% chloroauric acid solution, and make It was heated and boiled for 10 minutes and then cooled to 1 L for use.

[0072] Membrane drawing: take goat anti-mouse, urine microalbumin antigen (purchased from HYTEST), β2 microglobulin antigen (purchased from Guilin Inmate Biology), and respectively use 0.01mol / L PB+3% trehalose to prepare goat anti-mouse , urine microalbumin antigen and β2 microglobulin antigen were diluted to 1mg / mL, 3mg / mL and 1mg / mL successively, and the dosage was 35μL / 30cm, and the film was drawn onto a nitrocellulose membrane (Sartorius 2cm, 140 film), react at 42°C for 2h.

[0073] Marking: Take 5mL of colloidal gold solution and mix it with 30μL of 0.2mol / L potassium carbonate solution, take 30μg of urine ...

Embodiment 2

[0091]Dilute urine microalbumin antigen and β2 microglobulin antigen to 0.1ng / ml, 0.5ng / ml, 2ng / ml, 20ng / ml, 200ng / ml, 1μg / ml, 2μg / ml, 10μg / ml, 20μg / ml ml, 50μg / ml, take 30 strips respectively, and use 3 strips for each condition to test the test strips of Example 1 and Comparative Examples 1-3. Add the antigen to the sample pad and react directly, and see the result after 5 minutes. The results are shown in Table 1:

[0092] Table 1 Test strip detection limit

[0093]

[0094] The cut off values ​​of urinary microalbumin (MAU) and β2 microglobulin (β2-MG) were 20μg / ml and 0.2μg / ml, respectively.

[0095] It can be seen from Table 1 that:

[0096] Example 1 The detection limits of the test strip for urinary microalbumin and β2 microglobulin are 20 μg / ml and 200 ng / ml, respectively, which are consistent with the cut off values.

[0097] Comparative Example 1 The detection limits of urinary microalbumin and β2 microglobulin were above 50 μg / ml and 1 μg / ml respectively,

...

Embodiment 3

[0102] Take 10 urine microalbumin and β2 microglobulin positive specimens, and 10 normal human specimens for detection, and use the test strip of Example 1 for detection. 5 minutes to see the result. The test results are shown in Table 2:

[0103] Table 2 Test results

[0104]

[0105] Table 2 shows that some of the samples positive for microalbumin in urine were positive for β-microglobulin, and some of the samples positive for microalbumin in urine were also positive for microalbumin in urine. In the negative samples, the test results of both are negative. The result is fully consistent with expectations, indicating that the test strip provided by the present invention has a high accuracy rate.

[0106] Among them, positive results such as figure 1 , negative results such as figure 2 .

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Abstract

The invention relates to the technical field of biochemical detection, in particular to a test strip, a preparation method thereof and an application of the test strip to combined detection of microalbuminuria and beta2 microglobulin. According to the test strip, a sample pad, a gold mark pad, a chromatography membrane and absorbent paper are sequentially arranged on a bottom plate, and the gold mark pad and the chromatography membrane are coated with antibodies or antigens of specific content, so that detection limit of the test strip is effectively controlled and is a cut off value of albumin and the beta2 microglobulin. According to the test strip, the albumin and the beta2 microglobulin can be simultaneously detected by the aid of antibody concentration, and mutual interference is avoided. By the aid of the test strip, urine samples are directly detected, a pretreatment step is omitted, and detection of the test strip is rapider and more reliable. Experiments show that the test strip is high in accuracy, and the preparation method of the test strip is quite simple.

Description

technical field [0001] The invention relates to the technical field of biochemical detection, in particular to a test strip and its preparation method and its application in the joint detection of urinary microalbumin and β2 microglobulin. Background technique [0002] Microalbuminuria (MAU) refers to the presence of trace amounts of albumin in urine. Albumin is a normal protein in the blood. Under normal human metabolism, there is very little albumin in the urine, and the albumin in each liter of urine does not exceed 20μg (<20μg / mL). When renal vascular disease occurs, the function of the kidney to filter protein (especially albumin) is changed, and the amount of albumin in the urine will rise to more than 20 μg / mL. Therefore, clinically, urine microalbumin is usually used to monitor the occurrence of nephropathy, and it is used as an important indicator for detecting diabetes, hypertension, cardiovascular disease, and vascular damage in nephropathy. [0003] β2 micro...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893
Inventor 刘伟齐晶
Owner SINOCARE
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