Serum CENPF antibody (IgG and IgM) detection colloidal gold test strip

A technology for colloidal gold test strips and test strips, which is applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of inconvenient early case screening, long detection period, strong non-specific results, etc., and achieve intuitive and visual results. The effect of convenient and fast detection process

Pending Publication Date: 2021-06-01
BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In addition, in the prior art, indirect ELISA is often used to detect centromere protein F (CENPF) autoantibodies. Not only is the detection process complicated and the detection period is long, but the results are often non-specific and poor in repeatability. If it is used for screening a large number of clinical samples When testing, the workload is huge, the efficiency is low, and it is not convenient for large-scale early case screening

Method used

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  • Serum CENPF antibody (IgG and IgM) detection colloidal gold test strip
  • Serum CENPF antibody (IgG and IgM) detection colloidal gold test strip
  • Serum CENPF antibody (IgG and IgM) detection colloidal gold test strip

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Embodiment 1

[0023] The preparation of embodiment 1 colloidal gold test strip

[0024] 1. Materials

[0025] The CENPF protein and rabbit antibody used for the gold label antigen were prepared in our laboratory; mouse monoclonal antibody, goat anti-mouse secondary antibody for quality control line coating antibody, anti-human IgM antibody for T1 detection line, anti-human IgG antibody for T2 detection line, colloidal gold The solution, blocking solution, diluent, and gold complex solution were purchased from Nanjing Zhongding Biotechnology Co., Ltd.; absorbent paper, bottom plate, gold standard release pad and sample chromatography pad in the glass fiber pad were purchased from Millipore; nitrocellulose membrane Available from Sartorius Corporation.

[0026] 2. Preparation of colloidal gold test strips

[0027] 1. Labeled protein

[0028] Take two 1.5mL centrifuge tubes and wash them twice with ultrapure water, absorb 1mL colloidal gold and add to each tube, add 8μL 0.2M potassium carbo...

Embodiment 2

[0035] Example 2 Preparation of Rabbit Antibody for Detection Sensitivity

[0036] 1. Five New Zealand white rabbits were selected as experimental animals.

[0037] 2. Immunization: Inject Freund's complete adjuvant for initial immunization, and then except for the final immunization, all injections are injected with Freund's incomplete adjuvant for immunization. The immunogen and adjuvant are mixed, and the stable emulsion formed after thorough mixing is used. Spot injection for immunization.

[0038] 3. Serum collection and detection: After the animal immunization is completed, a small amount of serum can be used for titer detection by ELISA (results see figure 2 and Table 1). The titer is qualified for final blood collection.

[0039] 4. Purification of polyclonal antibodies: Antiserum was purified using Protein G and octanoic acid-ammonium sulfate.

[0040] Table 1. Determination of serum titer results of rabbit anti-CENPF ELISA method for detection sensitivity

[00...

Embodiment 3

[0042] Embodiment 3 carries out the detection of clinical sample with colloidal gold test strip

[0043] (1) Serum dilution

[0044] Use the colloidal gold test strip prepared by the present invention to detect the HCC serum sample, get every kind of serum sample 20uL and add 70uL diluent to dilute.

[0045] (2) Sample addition

[0046] Add the diluted sample dropwise into the sample hole of the test strip, and react for 10 minutes to interpret the result (see image 3 ).

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Abstract

The invention relates to a colloidal gold test strip which can be used for qualitative or semi-quantitative detection of two serum CENPF autoantibodies (IgG antibody and IgM antibody) at the same time. The detection test strip can rapidly qualitatively or semi-quantitatively detect the content of the CENPF antibody in serum through colloidal gold reaction, and can be used for rapid screening of early hepatocellular carcinoma. The colloidal gold test strip provides a novel hepatocellular carcinoma CENPF autoantibody detection method, the CENPF antibody detection sensitivity is about 4.1 ng / mL, the detection time is 10-15 minutes, the operation is simple and easy to implement, the result is visual and visible, and the colloidal gold test strip is suitable for rapid screening of early hepatocellular carcinoma.

Description

technical field [0001] The invention relates to a colloidal gold test strip for detecting serum CENPF antibodies (IgG and IgM). Background technique [0002] Hepatocellular Carcinoma (HCC) is the main type of primary liver cancer. The morbidity and mortality of liver cancer rank No. 6 and No. 4 in the incidence and death of cancers in the world. Due to the lack of early symptoms of HCC, most patients are already in the middle and late stage when symptoms appear, and they lose the chance of effective treatment. After HCC is found through early diagnosis, resection or liver transplantation is performed, and the 5-year survival rate of patients is usually above 60-70%. Therefore, early diagnosis can win time for surgical treatment for patients in clinic, and plays an irreplaceable role in improving survival rate and prognosis. However, at present, there is still a lack of HCC screening and early diagnosis methods with high sensitivity, good specificity, and clinical operabili...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/564G01N33/577G01N33/58G01N33/558
CPCG01N33/57438G01N33/57488G01N33/564G01N33/577G01N33/587G01N33/558G01N2333/47
Inventor 黄坚齐赛平陈质斌
Owner BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV
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