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Immunoblotting kit for detecting autoimmune nephrosis and preparation method thereof

An autoimmunity and immunoblotting technology, applied in the field of biomedicine, can solve problems such as delayed treatment and misdiagnosis, achieve intuitive and accurate result judgment, improve detection efficiency and accuracy of result judgment

Inactive Publication Date: 2016-08-03
SHENZHEN YHLO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the gold standard for the diagnosis and differential diagnosis of MN is still histopathology. IMN often has some histopathological features and can be differentiated from SMN, but sometimes it is difficult to distinguish the two completely even from renal histopathology difference, leading to misdiagnosis and delayed treatment

Method used

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  • Immunoblotting kit for detecting autoimmune nephrosis and preparation method thereof
  • Immunoblotting kit for detecting autoimmune nephrosis and preparation method thereof
  • Immunoblotting kit for detecting autoimmune nephrosis and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] C1q, MPO, PR3, GBM, PLA2R, THSD7A, Nuclesome and dsDNA antigens at a concentration of 0.05~0.1mg / mL, and human IgG, anti-mouse IgG or anti-goat IgG quality control at a concentration range of 30~50μg / mL The high-concentration indicator strip, the medium-concentration indicator strip with a concentration range of 12-25 μg / mL, and the low-concentration indicator strip with a concentration range of 1-10 μg / mL were prepared to the corresponding coating concentrations for use. According to the size of the immunoblotting spotter, plant the nitrocellulose membrane or nylon membrane into a membrane, and coat the prepared sample solution above in a certain order on the membrane through physical adsorption and covalent bonding. , forming an independent detection line and judgment indicator band. Put the coated film in a 37°C constant temperature oven to dry for 2 hours, take it out and stick it on the front of the double-sided adhesive PVC board, and the back of the PVC board is ...

Embodiment 2

[0090] Use C1q, MPO, PR3, GBM, PLA2R, THSD7A and Nuclesome antigens at a concentration of 0.05~0.1mg / mL, and human IgG, anti-mouse IgG or anti-goat IgG quality control at a high concentration of 30~50μg / mL The indicator strips, medium concentration indicator strips with a concentration range of 12-25 μg / mL, and low concentration indicator strips with a concentration range of 1-10 μg / mL were prepared to the corresponding coating concentrations for use. According to the size of the immunoblotting spotter, plant the nitrocellulose membrane or nylon membrane into a membrane, and coat the prepared sample solution above in a certain order on the membrane through physical adsorption and covalent bonding. , forming an independent detection line and judgment indicator band. Put the coated film in a 37°C constant temperature oven to dry for 2 hours, take it out and stick it on the front of the double-sided adhesive PVC board, and the back of the PVC board is ready for use, and then cut ...

Embodiment 3

[0092] Use C1q, MPO, PR3, GBM, PLA2R, THSD7A and dsDNA antigens at a concentration of 0.05~0.1mg / mL, and human IgG, anti-mouse IgG or anti-goat IgG quality control at a high concentration of 30~50μg / mL The indicator strips, medium concentration indicator strips with a concentration range of 12-25 μg / mL, and low concentration indicator strips with a concentration range of 1-10 μg / mL were prepared to the corresponding coating concentrations for use. According to the size of the immunoblotting spotter, plant the nitrocellulose membrane or nylon membrane into a membrane, and coat the prepared sample solution above in a certain order on the membrane through physical adsorption and covalent bonding. , forming an independent detection line and judgment indicator band. Put the coated film in a 37°C constant temperature oven to dry for 2 hours, take it out and stick it on the front of the double-sided adhesive PVC board, and the back of the PVC board is ready for use, and then cut it i...

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Abstract

The invention relates to an immunoblotting kit for detecting autoimmune nephrosis and a preparation method thereof, which belong to the field of biomedicine. The immunoblotting kit comprises a reaction membrane strip, an enzyme conjugate, a chromogenic substrate, a washing solution, a sample diluent and a stopping solution; the reaction membrane strip is composed of a carrier and a cellulose nitrate membrane or a nylon membrane which are fixed on the carrier, the cellulose nitrate membrane or the nylon membrane contains parallel detection lines specifically combined and coated by 4-8 kinds of natural or recombinant antigens in C1q, MPO, PR3, GBM, Nucleosome, dsDNA, PLA2R and YHSD7A, a strip of determination indication strip with high concentration being 30-50 [mu]L / mL, a strip of the determination indication strip with medium concentration being 15-25 [mu]L / mL, and a strip of the determination indication strip with low concentration being 1-10 [mu]L / mL. The kit can overcome the disadvantages of detection sensitivity and specificity of a single autoantibody as well as tedious operation for individually detecting several kinds of disease-associated autoantibodies, and can greatly increase the detection efficiency and result determination accuracy.

Description

technical field [0001] The invention relates to an immunoblotting kit for detecting multiple autoimmune nephropathy-related antibodies, and also relates to a preparation method and a use method of the immunoblotting kit, belonging to the field of biomedicine. Background technique [0002] The main function of the human immune system is to identify and remove antigenic foreign bodies. Under normal circumstances, the body generally does not produce an immune response to its own tissue components, which is called immune tolerance. Once this tolerance is broken, autoimmune disease occurs. Autoimmune diseases are characterized by the formation of various autoantibodies in the serum and the damage of multiple organs throughout the body. The kidney is one of the most common and important organs involved. In various autoimmune kidney diseases, some autoantibodies are not only the marker antibodies of certain types of diseases, but also may be direct pathogenic antibodies, which ar...

Claims

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Application Information

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IPC IPC(8): G01N33/564G01N33/548G01N33/545
CPCG01N33/564G01N33/545G01N33/548G01N2800/347
Inventor 何林胡鹍辉刘清波阳辉
Owner SHENZHEN YHLO BIOTECH
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