Serum CENPF antibody quantitative detection kit

Abstract

The invention relates to a serum CENPF antibody quantitative detection kit which can be used for specifically and quantitatively detecting the level of a CENPF antibody in a serum specimen. According to the serum CENPF antibody quantitative detection kit, recombinant CENPF antigen protein is coated in a 96-pore enzyme label board, the level of the CENPF antibody in the serum specimen can be quantitatively tested by using an enzyme linked immunosorbent assay, and the disease state of a liver cell cancer patient or high risk groups can be evaluated according to the level of the CENPF antibody in the serum. By adopting the serum CENPF antibody quantitative detection kit, a novel liver cell cancer screening and early diagnosis method is provided, and the sensitivity of the detected serum CENPF self-antibody is remarkably superior to that of a conventional clinical serum marker AFP when being used for diagnosing liver cell cancer.

Description

technical field

[0001] The invention relates to a serum CENPF antibody quantitative detection kit. Background technique

[0002] Hepatocellular Carcinoma (HCC) is the main type of primary liver cancer. Due to the lack of early symptoms of HCC, most patients are already in the middle and late stages when symptoms appear, losing the chance of effective treatment, and the survival period is generally less than 1 year. Early HCC can be resected or liver transplanted, and the 5-year survival rate is above 60-70%. . Therefore, timely detection and diagnosis of early cases through regular screening of high-risk groups is crucial. However, at present, there is still a lack of HCC screening and early diagnosis methods with high sensitivity, good specificity, and clinical operability.

[0003] An ideal early diagnosis method for HCC should not only have high specificity to distinguish HCC from liver cirrhosis and hepatitis, but also have high sensitivity to detect HCC at an early s...

Examples

Embodiment 1

[0019] Example 1 Preparation of CENPF recombinant protein

[0020] 1. Materials

[0021] 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; GST antibody was purchased from Beyontien 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.

[0022] 2. Construction and identification of human CENPF prokaryoti...

Embodiment 2

[0035] Example 2 Preparation of ELISA plate

[0036] 1. Materials

[0037] The 96-well ELISA plate was purchased from Thermo Company in the United States; the recombinant CENPF antigen protein was prepared in our laboratory; BSA was purchased from Beijing Huanya Tech Biomedical Technology Co., Ltd.; TMB color development solution and stop solution were purchased from Beijing Solaibao Technology Co., Ltd. company; HRP-labeled rabbit anti-human IgG was purchased from Sigma, USA; other reagents were of domestic analytical grade.

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

[0039] In the previous research process, our research group has quantitatively detected the serum CENPF antibody level of a group of liver cancer patients through protein chip technology, and screened 30 cases of serum containing high levels of CENPF antibody (protein chip detection signal value greater than 30,000) The serum samples were mixed as the standard of this test kit.

[0040...

technical field

[0001] The invention relates to a serum CENPF antibody quantitative detection kit. Background technique

[0002] Hepatocellular Carcinoma (HCC) is the main type of primary liver cancer. Due to the lack of early symptoms of HCC, most patients are already in the middle and late stages when symptoms appear, losing the chance of effective treatment, and the survival period is generally less than 1 year. Early HCC can be resected or liver transplanted, and the 5-year survival rate is above 60-70%. . Therefore, timely detection and diagnosis of early cases through regular screening of high-risk groups is crucial. However, at present, there is still a lack of HCC screening and early diagnosis methods with high sensitivity, good specificity, and clinical operability.

[0003] An ideal early diagnosis method for HCC should not only have high specificity to distinguish HCC from liver cirrhosis and hepatitis, but also have high sensitivity to detect HCC at an early s...

Examples

Embodiment 1

[0019] Example 1 Preparation of CENPF recombinant protein

[0020] 1. Materials

[0021] 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; GST antibody was purchased from Beyontien 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.

[0022] 2. Construction and identification of human CENPF prokaryoti...

Embodiment 2

[0035] Example 2 Preparation of ELISA plate

[0036] 1. Materials

[0037] The 96-well ELISA plate was purchased from Thermo Company in the United States; the recombinant CENPF antigen protein was prepared in our laboratory; BSA was purchased from Beijing Huanya Tech Biomedical Technology Co., Ltd.; TMB color development solution and stop solution were purchased from Beijing Solaibao Technology Co., Ltd. company; HRP-labeled rabbit anti-human IgG was purchased from Sigma, USA; other reagents were of domestic analytical grade.

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

[0039] In the previous research process, our research group has quantitatively detected the serum CENPF antibody level of a group of liver cancer patients through protein chip technology, and screened 30 cases of serum containing high levels of CENPF antibody (protein chip detection signal value greater than 30,000) The serum samples were mixed as the standard of this test kit.

[0040...

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