Diagnostic marker for systemic lupus erythematosus

A lupus erythematosus, systematic technology, applied in the direction of material inspection products, measuring devices, instruments, etc., can solve problems such as easy missed diagnosis

Active Publication Date: 2015-05-27
杭州中赢生物医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this standard is easy to miss diagnosis of early and atypical cases, so a new diagnosti...

Method used

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  • Diagnostic marker for systemic lupus erythematosus
  • Diagnostic marker for systemic lupus erythematosus
  • Diagnostic marker for systemic lupus erythematosus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Using the PNGase F enzyme kit (IBL, Germany), according to the kit instructions provided by the manufacturer, the N-glycan chains of serum antibodies were extracted, and the N-glycan chains of serum antibodies were analyzed by primary and multi-stage mass spectrometry. The process is as follows:

[0034] (1) Collect serum samples from SLE patients and centrifuge at 2000r / min for 10min.

[0035] (2) Take the supernatant, dilute it at the ratio of 1mL serum to 3mL PBS buffer, add saturated ammonium sulfate buffer dropwise, let stand for 3h, and then centrifuge at 6000r / min.

[0036] (3) The precipitate obtained in step (2) was further desalted with sephadex G25, and then passed through a proteinG column to further purify the antibody, and freeze-dried to obtain serum antibody, which was stored at -20°C for future use.

[0037] (4) Put 500 micrograms of serum antibody into a centrifuge tube, add 5 microliters of 10× denaturant and 45 microliters of ultrapure water, mix wel...

Embodiment 2

[0058] Construct the SLE mouse model:

[0059] BALB / c mice aged 6-8 weeks were purchased and raised in an SPF animal room. After the environment was stabilized, they were randomly divided into two groups, with 5 mice in each group. The mice in the experimental group (ie, the SLE model group) were intraperitoneally injected with 0.5 mL of pristane (purchased from sigma), while the mice in the control group were injected with 0.5 mL of PBS. Blood was collected once a week before and after the injection, and ELISA and other methods were used to detect the contents of anti-dsDNA, anti-sm RNP and anti-ribosomal PO antibodies, and finally confirmed the success of the establishment of the SLE mouse model.

[0060] Treatment: Feed the mice for about 6 months, kill them, and take the serum for future use. According to the method in Example 1, N sugar chains were extracted, and after primary and multi-stage mass spectrometry detection, the primary mass spectrogram was as follows Figu...

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Abstract

The invention relates to a diagnostic marker for systemic lupus erythematosus, and in particular discloses application of a reagent for specific detection of high mannose type N-linked carbohydrate chain level in a serum antibody to preparation of a diagnostic tool for diagnosis and/or prognosis assessment of systemic lupus erythematosus. The increase of the high mannose type N-linked carbohydrate chain level in the serum antibody is closely related to the occurrence of systemic lupus erythematosus. Therefore, the diagnostic marker can be used for early diagnosis and/or prognostic assessment of systemic lupus erythematosus, and further can be used for guiding treatment of systemic lupus erythematosus.

Description

technical field [0001] The invention relates to the technical field of disease diagnosis, in particular to a diagnostic marker of systemic lupus erythematosus and its application. Background technique [0002] Systemic lupus erythematosus (SLE) is a diffuse, systemic autoimmune disease. It has a variety of clinical symptoms, involving skin and mucous membranes, skeletal muscles, kidneys and central nervous system diseases, and sometimes causes dysfunction of other organs and systems such as the lung and heart. SLE is more common in young women, with a peak incidence It is 15 to 40 years old, and the male to female ratio is about 1:9. Although studies have found that the onset tends to cluster in families, its pathogenesis is not a single genetic factor. The pathogenesis of systemic lupus erythematosus is still unclear. [0003] Studies have found that in the serum of patients with systemic lupus erythematosus, a variety of autoantibodies and immunological abnormalities can...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N30/72G01N30/06
CPCG01N33/577G01N33/6812
Inventor 吴忠福李云森
Owner 杭州中赢生物医疗科技有限公司
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