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A method for detecting complex Iga1-α1 mg and a kit for non-invasive detection of renal tissue damage

A kidney tissue and kit technology, applied in the field of IgA nephropathy detection methods and kits, can solve the problems of non-invasive detection of IgA nephropathy, and achieve the effect of low detection equipment requirements, good specificity, and small error

Active Publication Date: 2021-03-09
PEKING UNIV FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the technical problem of being unable to non-invasively detect IgA nephropathy in this field, the inventor found through extensive research that the complex IgA1-α1MG is the key pathogenic factor of IgA nephropathy. Therefore, the present invention detects the complex IgA1 in the sample - The detection of α1MG realizes the non-invasive detection of IgA nephropathy and renal tissue damage. The detection method and kit provided by the present invention can not only accurately quantify the concentration level of the pathogenic factor complex IgA1-α1MG in the circulation, but also timely and effectively detect Whether there is damage to the patient's kidney tissue and will not cause trauma to the patient

Method used

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  • A method for detecting complex Iga1-α1 mg and a kit for non-invasive detection of renal tissue damage
  • A method for detecting complex Iga1-α1 mg and a kit for non-invasive detection of renal tissue damage
  • A method for detecting complex Iga1-α1 mg and a kit for non-invasive detection of renal tissue damage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 The method for detecting complex IgA1-α1MG

[0065] 1. Package board

[0066] Dilute the unlabeled anti-human α1MG antibody to 5ug / ml with 0.05M carbonate buffer, 100ul per well, coat the plate, immobilize the anti-human α1MG antibody on the solid phase carrier, and incubate overnight at 4°C ; Then use 0.05% PBST, 360ul per well to wash the plate, after washing 5 times, the free components that are not immobilized on the solid phase carrier are washed away, use 1% BSA-PBST to block, 200ul per well, incubate at 25°C for 2h, Finally, the plate was washed with 0.05% PBST, 360ul per well, and washed 5 times to obtain a solid phase carrier coated with anti-human α1MG antibody.

[0067] Wherein, the solid phase carrier used in the present invention can be any commercially available solid phase carrier with good adsorption performance, low blank space, high transparency at the bottom of the hole, and similar performance between each plate and between each well of th...

Embodiment 2

[0079] Example 2 Kit for non-invasive detection of renal tissue damage

[0080] Test kit of the present invention comprises:

[0081] 1. A capture system for capturing IgA1-α1MG complex in plasma, including a solid carrier coated with anti-human α1MG, 0.05M carbonate buffer diluent, 0.05% PBST washing solution and 1% BSA-PBST blocking solution.

[0082] Among them, 0.05% PBST solution includes 8.0g NaCl, 2.9g Na 2 HPO4, 0.2g KCl, 0.26g KH 2 PO 4 , 1000ml H 2 O and 0.5ml Tween-20 mixed, the pH value is 7.4.

[0083] Among them, 0.05M carbonate buffer contains 1.08gNa 2 CO 3 , 2.95g NaHCO 3 , 900ml H 2 O mix, pH 9.6.

[0084] Wherein, the 1% BSA-PBST solution includes 1 g of BSA and 100 ml of 0.05% PBST solution.

[0085] 2. The detection system for detecting IgA1-α1MG complex, including

[0086] 1% BSA-PBST solution, used to dilute test samples and standards, or to dilute avidin ExtraAvidin, 0.05% PBST washing solution, biotin biotin and avidin Avidin reaction system...

Embodiment 3

[0088] Example 3 Kit for non-invasive detection of renal tissue damage

[0089] Including 1 and 2 in embodiment 3, and a judging system for determining whether the sample is a kidney tissue damage sample.

[0090] Further, the judging system used to determine whether the sample is a kidney tissue damage sample is as follows: figure 2 As shown, it includes: a data reading unit 1 connected with a microplate reader, which is used to directly read the OD value detected by the microplate reader, and obtain the detection value of the sample to be tested and the detection value of the standard sample; and the data reading unit 1 The connected data analysis calculation unit 2 is used to form the standard curve with the OD value of the measured standard sample, and calculate the concentration level of the detection value of the sample to be tested according to the formed standard curve; the data judgment unit connected with the data analysis calculation unit 2 3. Comparing the concen...

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Abstract

The invention provides a method for detecting complex IgA1-α1MG and a kit for noninvasively detecting renal tissue damage, which relate to the field of biotechnology, comprising: using an anti-human α1MG antibody to capture the complex IgA1-α1MG in a sample to be tested to obtain the test kit Complex IgA1-α1MG; adding a marker-labeled anti-human IgA1 antibody to the complex IgA1-α1MG to be tested, so that the content of the anti-human IgA1 antibody labeled with a marker is the same as the content of the complex IgA1-α1MG to be tested It is positively correlated; by detecting the marker of the anti-human IgA1 antibody to obtain the detection result of whether there is a complex IgA1-α1MG in the sample to be tested, the present invention realizes the detection of IgA nephropathy, which fills the gap in the detection of IgA nephropathy in the technical field Blank, breaking the technical problem in the technical field that the tissue damage of IgA nephropathy cannot be detected non-invasively.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a detection method and kit for IgA nephropathy. Background technique [0002] IgA nephropathy is the most common primary glomerulonephritis worldwide. In my country, IgA nephropathy accounts for as high as 58.2% of the cases of primary glomerular disease in renal biopsy. The most commonly affected population of IgA nephropathy is young and middle-aged males; and the course of the disease progresses slowly, 25% to 30% of IgA nephropathy patients progress to requiring renal replacement therapy within 20 to 25 years after onset, causing a huge disease burden to the society. At present, kidney biopsy, an invasive examination, is generally used for the detection of nephropathy, which is more harmful to patients. [0003] The characteristic manifestation of IgA nephropathy is the mass deposition of IgA-based immunoglobulin molecules in the glomerular mesangium. Clinical observations hav...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/6854G01N33/6893G01N2800/347
Inventor 张宏朱厉
Owner PEKING UNIV FIRST HOSPITAL
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