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Serum ALDH1B1 autoantibody quantitative detection kit and application thereof

A technology for quantitative detection of autoantibodies, applied in the field of protein marker detection, can solve problems such as lack of research

Active Publication Date: 2020-10-02
BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the reported markers are still lacking in early colorectal cancer and high-grade adenoma (precancerous lesions).

Method used

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  • Serum ALDH1B1 autoantibody quantitative detection kit and application thereof
  • Serum ALDH1B1 autoantibody quantitative detection kit and application thereof
  • Serum ALDH1B1 autoantibody quantitative detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The preparation of embodiment 1 recombinant ALDH1B1 antigenic protein

[0026] 1. Materials

[0027]DNA marker (100-2000bp) and rainbow prestained protein marker (14-120KD) were purchased from Beijing Tiangen Biochemical Technology Co., Ltd.; chemiluminescent protein marker (20-90KD) was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; Bacillus Origammi2 and BL21 (DE3) were preserved in our laboratory; Fast Taq Mastermix, BamHI, XhoI, and T4 DNA ligase were purchased from NEB Company; DNA electrophoresis gel recovery kit was purchased from Axygen Company; D-galactoside (IPTG) was purchased from Amresco; ALDH1B1 antibody was purchased from Beyond Biotechnology Research Institute; HRP-labeled goat anti-human IgG was purchased from Earthox, USA; primer synthesis and DNA sequence determination were provided by Shanghai Sangon Bioengineering Co., Ltd. Finish. All other reagents were of domestic analytical grade.

[0028] 2. Construction and identification of hum...

Embodiment 2

[0035] The preparation of embodiment 2 standard substance and microtiter plate

[0036] 1. Materials

[0037] The 96-well ELISA plate was purchased from Thermo Company in the United States; the recombinant ALDH1B1 antigen protein was prepared according to the method in Example 1; BSA was purchased from Beijing Huanya Tech Biomedical Technology Co., Ltd.; TMB chromogenic solution and stop solution were purchased from Beijing Solai Bao Technology Co., Ltd.; HRP-labeled rabbit anti-human IgG was purchased from Sigma, USA; other reagents were domestic analytical grade.

[0038] 2. Preparation of standard products in the detection kit

[0039] The rabbit polyclonal antibody against human ALDH1B1 was obtained by immunizing rabbits with the recombinant ALDH1B1 antigen protein prepared in Example 1, and the obtained serum antibody was further purified by affinity chromatography and used as the quantitative standard of the kit. In addition, referring to the method of Zhang Jinlong et...

Embodiment 3

[0054] Example 3 Diagnostic value of ALDH1B1 autoantibodies in colorectal cancer and colorectal high-grade adenoma

[0055] ALDH1B1 autoantibody was detected on 315 samples, including 110 healthy samples, 75 colorectal high-grade adenomas and 130 colorectal cancer serum samples, using the kit method, and the diagnostic value was evaluated by ROC curve and AUC value.

[0056] The results showed that the AUC values ​​of ALDH1B1 autoantibody in colorectal high-grade adenoma and colorectal cancer were 0.74 and 0.70, and the sensitivity and specificity were 75.68%, 62.31%, 63.06%, and 73.87%, respectively; The positive rate of early colorectal cancer was only 38.6%, and the positive rate of ALDH1B1 was 62.3%. More than half of CEA-negative early colorectal cancer were positive for ALDH1B1 autoantibody. Experimental results such as image 3 and Figure 4 shown.

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Abstract

The invention discloses a serum ALDH1B1 autoantibody quantitative detection kit and an application thereof. The kit comprises a solid-phase carrier, a recombinant ALDH1B1 antigen protein coated on thesolid-phase carrier, an enzyme-labeled secondary antibody, a standard substance, a chromogenic substrate, a washing solution and a reaction stop solution. The kit can be used for specifically and quantitatively detecting the level of the ALDH1B1 antibody in a serum sample, and is obviously superior to a serum marker CEA which is clinically used at present in the aspects of early screening and diagnosis of colorectal cancer and high-grade colorectal adenoma sensitivity.

Description

technical field [0001] The invention belongs to the field of protein marker detection, in particular to a detection kit for early colorectal cancer and high-grade colorectal adenoma based on the detection of serum ALDH1B1 autoantibody. Background technique [0002] Cancer is the leading cause of death, and a large number of cancer cases are diagnosed every year. According to global cancer estimates, there were 18.1 million new cancer cases and 9.6 million deaths in 2018, including 1.8 million new cases of colorectal cancer and 881,000 deaths. Among them, new cancer cases in China accounted for 24% (4.3 million), cancer deaths accounted for 30% (2.9 million), and colorectal cancer accounted for 12.8% of malignant tumors. Cancer has become the leading cause of death in China. The cancer death rate in the United States dropped from a peak of 215.1 / 100,000 in 1991 to 158.6 in 2015, a decline rate of 26%. Specifically, from 1970 to 2015, the death rate from colorectal cancer d...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/564G01N33/543
CPCG01N33/57419G01N33/57488G01N33/564G01N33/54393G01N2333/90203Y02A50/30
Inventor 黄坚王何静张蓓李潇瑾
Owner BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV
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