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Kit and detection method for determining anti-Jo-1 antibody

A kit and reagent technology, applied in the field of kits for the determination of anti-Jo-1 antibodies, can solve the problems of cross-contamination of reagents, complicated operations, affecting test results, etc., and achieve the improvement of positive signal value, simple and convenient operation, and avoid result deviation. Effect

Active Publication Date: 2018-12-25
SUZHOU HAOOUBO BIOPHARML
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] (1) False positives may occur using this method
[0008] (2) Non-specific reactions cannot be distinguished according to the size of the molecular weight when analyzing the results
[0009] (3) The operation is relatively complicated and requires expensive fluorescence microscopes, which are difficult to promote in many primary hospitals. At the same time, it is not suitable for laboratories and diagnostic hospitals with large sample volumes
[0010] (4) The background in fluorescence measurement is high, and it is difficult to use fluorescence immunoassay for quantitative determination
[0011] (5) Experienced professionals are required to judge the results, and the objectivity of the analysis results is insufficient
[0014] (1) The detection reagent is an open method during the detection process, which is not only easily affected by the external environment, but also easily causes cross-contamination between various reagents and affects the detection results;
[0015] (2) ELISA detection range and sensitivity are relatively low
[0016] (3) The detection time of the ELISA method is long, and it generally takes more than 2 hours to complete a test, which cannot fully meet the needs of clinical rapid diagnosis
[0017] (4) The ELISA method cannot be randomly injected for detection, and there is a lag in the detection results
Only biotin-labeled Jo-1 antigen is used in this patent, which makes the detection sensitivity of anti-Jo-1 antibody lower

Method used

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  • Kit and detection method for determining anti-Jo-1 antibody

Examples

Experimental program
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Effect test

Embodiment 1

[0057] (1) Preparation of the first reagent

[0058] 1. Materials and instruments:

[0059] Materials: Jo-1 antigen, biotin activated by N-hydroxysuccinimide, tris buffer, glycerol, phosphate buffer;

[0060] Instruments: Reagent cryogenic storage box, biochemical incubator.

[0061] 2. Preparation steps:

[0062] Step 1: Mix 2mg of Jo-1 antigen with 0.5mg of biotin activated by N-hydroxysuccinimide, mix and react at 25°C for 30min;

[0063] Step 2: Add 20uL of tris buffer solution with a substance concentration of 0.05mol / L, mix and react at 30°C for 30min, then add 600uL of glycerol to obtain biotinylated Jo-1 antigen. Store at -20°C for later use;

[0064] Step 3: Dilute the biotinylated Jo-1 antigen with a phosphate buffer solution with a pH of 7.5 and a substance concentration of 0.01 mol / L to a mixed solution with a concentration of 1 ug / mL.

[0065] Step 4: Add histidine tRNA with a final concentration of 0.6ug / mL and an RNase inhibitor with a final mass concentrat...

Embodiment 2

[0096] Embodiment 2: Fully automatic chemiluminescence measuring instrument test procedure

[0097] The automatic chemiluminescence analyzer test proceeds in the following steps:

[0098] Step 1: The detection kit needs to be used in conjunction with the automatic chemiluminescence analyzer of our company. Put the kit into the corresponding position of the reagent compartment of the automatic chemiluminescence analyzer, and input the information of the kit into the instrument system through the barcode scanner or through the supporting software of the instrument set up.

[0099] Step 2: Place the calibrator in the sample compartment of the instrument. Identify the calibrator information through the barcode scanner and assign the calibrator location in the instrument system.

[0100] Step 3: Place the quality control substance / sample to be tested in the sample compartment of the instrument, and edit the corresponding detection information through the supporting software of th...

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Abstract

The invention relates to a kit for determining an anti-Jo-1 antibody. The kit comprises a first reagent, wherein the first reagent comprises a mixture coupled with biotin. The mixture comprises a Jo-1antigen and a tRNA. According to the kit and the detection method for determining anti-Jo-1 antibody, the invention has the advantages of good stability, high sensitivity, good repeatability and thelike; a detection system can realize random sample loading, freely combination and continuously detecting, and the detection time is greatly reduced that 120 tests can be completed within 1 hour; andoperations are simple and convenient, a full automatic detection is truly realized, and the result deviation caused by the operations is avoided. Compared with a traditional method, the detection result of the invention is consistent in conformity, and the detection rate of a Jo-1 antibody positive in dermatomyositis / polymyositis is increased to 35%-40%, furthermore the positive signal value is increased by 10-20%.

Description

technical field [0001] The invention belongs to the technical field of magnetic particle chemiluminescence immunodiagnosis, and in particular relates to a kit and a detection method for measuring anti-Jo-1 antibody. Background technique [0002] Histidyl-tRNA synthetase (Jo-1) is a cytoplasmic phosphoprotein with a molecular weight of 50kDa, which can connect the free histidine in the cytoplasm to the corresponding tRNA and convert it into mRNA Plays an important role in forming peptides as templates. At the same time, we found that some Jo-1 antibody positive patient sera can react with tRNA. [0003] Anti-Jo-1 antibody is common in polymyositis (Polymyositis; PM), and the positive detection rate can reach 40%-50%, and the positive detection rate in PM / DM (dermatomyositis) patients is 25%. The detection rate is less than 10% in other autoimmune diseases, anti-Jo-1 antibody is negative, so anti-Jo-1 antibody is specific for the diagnosis of PM. In patients with PM overlap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543
CPCG01N33/535G01N33/54326
Inventor 杨苏清柳乐崔利歌黎静雯赵婷郅晓乐孔巧芸喻露郝佳丽李波李裕明李庆春
Owner SUZHOU HAOOUBO BIOPHARML
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