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Detection material resisting NMDAR autoantibodies in human body fluid, preparation method and application

A technology of autoantibodies and detection materials, applied in the field of biomedicine, can solve the problems of large demand for antibodies, long detection time, low sensitivity, etc., and achieve the effect of strong operation skills, short detection cycle and high detection cost

Inactive Publication Date: 2019-12-24
SHAANXI MYBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problems existing in the prior art, the purpose of the present invention is to provide a detection material for detecting NMDAR autoantibodies and the preparation method and application of the material, so as to solve the problem of time-consuming detection, cumbersome steps and antibody demand of existing detection methods. Large size, high cost, low sensitivity, etc.

Method used

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  • Detection material resisting NMDAR autoantibodies in human body fluid, preparation method and application
  • Detection material resisting NMDAR autoantibodies in human body fluid, preparation method and application
  • Detection material resisting NMDAR autoantibodies in human body fluid, preparation method and application

Examples

Experimental program
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Embodiment 1

[0038] Step 1, plasmid construction: Obtain the CDS sequence of NMDAR as the target gene, insert the target gene with restriction sites into the pShuttle-cmv plasmid vector to obtain the recombinant plasmid vector pAdEasy-NMDAR1α, and extract the plasmid for subsequent experiments after the sequencing is correct. The plasmid vector construction of the present embodiment specifically includes the following steps:

[0039]Step 1.1: Obtain the CDS sequence of NMDAR as the target gene by PCR method (artificial synthesis method is also optional), and add SalI / NotI restriction sites at both ends of the target gene;

[0040] Step 1.2: Insert the target gene with the restriction site into the pShuttle-cmv vector, the insertion site is SalI / NotI, to obtain the recombinant vector, which is named pShuttle-NMDAR1α;

[0041] Step 1.3: Sequence the recombinant plasmid pShuttle-NMDAR1α, expand and culture the correctly sequenced strains with the target gene and carrier strains (auxiliary vec...

Embodiment 2

[0063] The difference between this example and Example 1 is that the composite extract in step 3.1 is a mixture of digitonin, TritonX100, NP40, and CHAPS in a volume ratio of 1:1:1:1.

Embodiment 3

[0065] The difference between this example and Example 1 is that the composite extract in step 3.1 is a mixture of digitonin, TritonX100, NP40, and CHAPS in a volume ratio of 1:2:2:1.

[0066] When the anti-NMDAR autoantibody detection material prepared in the above examples is used to detect the NMDAR antibody in the sample, the present invention firstly prepares a new blocking material. Since some proteins in human serum and cerebrospinal fluid will non-specifically bind to antigens during the detection process and interfere with the detection results, the present invention uses a sealing material to seal the detection sample, and the implementation effect shows that the sample background can be significantly reduced, as follows A specific example of the preparation of the sealing material of the present invention is given.

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Abstract

The invention discloses a detection material resisting NMDAR autoantibodies in human body fluid and a preparation method and application thereof. According to the invention, NMDAR antigens are arranged on a carrier membrane through coating to prepare an anti-NMDAR-receptor detection material, specific NMDAR antibodies in human serum and cerebrospinal fluid can be combined with the antigens, alkaline phosphatase substrate-ligand reaction is utilized for color development, the sealing material is added in the color development reaction, and whether a detected sample contains the NMDAR antibodiesor not can be directly judged through naked eye observation. The method has the advantages of high sensitivity, easy and convenient operation and rapid detection, and is conducive to recognition anddiagnosis of anti-NMDAR-receptor encephalitis.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a detection material, preparation method and application of anti-NMDAR autoantibody in human body fluid. Background technique [0002] Anti-N-methyl-D-aspartate receptor (N-methyl-D-aspartate receptor, NMDAR) limbic encephalitis (hereinafter referred to as anti-NMDAR encephalitis) is an autoimmune disease, the first case in the world was in 2007 It was discovered by RobertaVitaliani et al. in 1999. In the same year, Josep Dalmau et al. detected anti-NMDAR antibodies in the serum and cerebrospinal fluid of such patients, and used this as the main basis for diagnosing this type of disease. [0003] The current diagnostic methods for anti-NMDAR encephalitis include medical history inquiry, general medical examination, X-ray and ultrasound, neuroimaging, blood test and cerebrospinal fluid examination, among which the positive NMDAR-specific antibody in blood and cereb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/705C07K1/14C12N15/861G01N33/531G01N33/564
CPCC07K14/70571C12N15/86C12N2710/10343C12N2800/107G01N33/531G01N33/564G01N2333/70571G01N2800/28
Inventor 闫亚平李怡婷郝文斌李科
Owner SHAANXI MYBIOTECH CO LTD
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