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Kit for combined detection of 6 diabetic antibodies

A technology of autoantibodies and diabetes, applied in the biological field, to achieve the effect of ensuring accuracy, simple operation, and improving detection efficiency

Active Publication Date: 2013-03-13
SHENZHEN BLOT BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there has not been any report on the use of western blotting in the six joint tests of diabetes autoantibodies in China.

Method used

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  • Kit for combined detection of 6 diabetic antibodies

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The preparation of embodiment 1 kit

[0049] Extraction and purification of 6 islet cell protein antigens including IA, GAD, IA-2, CPH, ZnT8 and ICA:

[0050] Screen pancreatic tissues from pigs, cattle, rats and humans, and select pancreatic tissues containing 6 islet cell protein antigens;

[0051] Extract 6 kinds of islet cell protein antigens: crush the pancreatic tissue with an ultrasonic cell breaker, put it in a high-speed refrigerated centrifuge, collect the supernatant, put it in a dialysis bag for dialysis, concentrate it in polyethylene glycol, and measure DM with an ultraviolet spectrophotometer protein antigen content. The key problem it mainly solves is that in the process of extracting and purifying DM antigen, its antigenic components will be degraded and lost.

[0052] Electrophoresis: The six antigens of islet cells were properly diluted with sample buffer, and SDS-polyacrylamide gel gradient discontinuous electrophoresis was used to separate various...

Embodiment 2

[0063] Embodiment 2 is based on the detection method of the kit provided by the invention

[0064] Serum or whole blood to be tested is 10 μl. Fresh serum or whole blood is suitable for the specimen to be tested. Serum can generally be collected from veins and centrifuged after coagulation. Point-of-care testing can be performed by finger pricking and whole blood testing. If the collected serum or whole blood is not tested on the same day, it should be stored at 4°C; if it exceeds seven days, it should be stored below -20°C.

[0065] Preparation of washing solution: the concentrated washing solution stored at 2-8°C will crystallize, dilute the whole bottle (25mL) of concentrated washing solution to 250mL with distilled water or pure water, put it into the reagent bottle, and indicate the preparation on the label Time, stored at 2-8°C, valid for 3 months, if there is floc or mildew, it should be discarded;

[0066] Use tweezers to put the required amount of blotting membrane ...

Embodiment 3

[0075] Comparison of several detection methods of embodiment 3

[0076] 45 cases of T1DM patients were newly diagnosed patients from the Department of Secretology and Pediatrics, all of which met the 1999 diagnostic criteria of the World Health Organization (WHO). Aged 5-48 years old, average age (25±12 years old), including 27 males and 18 females, healthy control group was 50 cases who came to our hospital for physical examination, aged 15-41 years old, average age (24±5 years old) , including 24 males and 26 females, all the controls had normal fasting blood sugar, no family history of diabetes and autoimmune disease history.

[0077]All tested samples were fasting venous blood collection, separated serum, aliquoted and stored at -20°C for testing. The IAA radioimmunoassay kit was purchased from the Beijing Institute of Biotechnology, and the SN-697 fully automatic dual-probe radioimmuno r-counter was produced by the Shanghai Atomic Energy Research Institute Rihuan Instrum...

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Abstract

The invention relates to the technical field of biology, and in particular relates to a kit for combined detection of 6 diabetic antibodies. The kit is used for combined detection of 6 antibodies such as ICA, GADA, IAA, IA-2ACPH-A and ZnT8-A in T1DM, T2DM and serum of a normal person, has high sensitivity (98 percent) and specificity (99.6 percent) for I type diabetic diagnosis, and provides efficient and fast detection means for classificatory diagnosis of diabetes.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a six-item joint detection kit for diabetes autoantibodies. Background technique [0002] Diabetes (diabetes) is a sugar, protein, A series of metabolic disorder syndromes including fat, water and electrolytes, clinically characterized by hyperglycemia, polyuria, polydipsia, polyphagia, weight loss, etc. Blood sugar) Once it is not well controlled, it will cause complications, leading to failure and lesions of the kidneys, eyes, feet and other parts, and it cannot be cured. Diabetes has become the third major disease that endangers human health after cardiovascular and cerebrovascular diseases and cancer. [0003] According to the etiology and pathogenesis clinically, it is divided into type 1 diabetes mellitus (T 1 DM), type 2 diabetes (T 2 DM) and other types. T 1 DM, also known as immune-mediated diabetes, is an autoimmune disease in which the synthesis and secretion of insu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/531
Inventor 马伟民张永顶马新民
Owner SHENZHEN BLOT BIOTECH
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