A method for rapid intraoperative identification of cervical lymph node metastasis in papillary thyroid carcinoma

A technology for lymph node metastasis and papillary carcinoma, applied in instruments, measuring devices, scientific instruments, etc., can solve the problems of low contrast between color strips and background, inability to realize quantitative detection of test objects, and difficulty in realizing multiple inspections and joint inspections, etc. To achieve the effect of reducing the incidence of hypoparathyroidism, reducing external conditions and background, and shortening the length of hospital stay

Active Publication Date: 2022-07-01
JIANGSU INST OF NUCLEAR MEDICINE
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0006] (1) The colloidal gold labeling process is an electrostatic adsorption process, which is a physical adsorption, so the stability in the liquid phase is poor, often causing the labeled protein molecules to fall off again
[0007] (2) The test result is judged by displaying a single purple-red strip, and the color is single, so it is difficult to realize multiple inspection and joint inspection
[0008] (3) Only when the gold particles gather to a certain amount, the human naked eye can observe the purple-red band, and the color band has little contrast with the background, thus limiting the detection sensitivity
[0009] (4) The matrix effect of different materials is obvious, and the background interference is very large
[0010] (5) The detection sensitivity is low
[0011] (6) Quantitative detection cannot be realized
[0012] At present, there is also a detection method that uses quantum dots to label rapid immunochromatographic test strips, but this method cannot achieve quantitative detection of the test substance.
[0013] In addition, in the immunoassay methods for detecting Tg levels, there is still a lack of combinations that can be detected with high sensitivity and high specificity in double-antibody sandwich assays. Finding more suitable immunogenic Tg epitope peptides, preparing specific Sexual Tg antigens and antibodies are also the key points to be solved

Method used

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  • A method for rapid intraoperative identification of cervical lymph node metastasis in papillary thyroid carcinoma
  • A method for rapid intraoperative identification of cervical lymph node metastasis in papillary thyroid carcinoma
  • A method for rapid intraoperative identification of cervical lymph node metastasis in papillary thyroid carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Screening of human Tg antigen peptides

[0046] The Tg described in this application is known in the art, and a complete Tg is composed of a single polypeptide chain containing 2768 amino acids, with a molecular weight of about 660,000 Daltons. Its amino acid sequence is known in the art and can be found in professional databases such as NCBI, and the specific sequence is as follows:

[0047] Human Tg(1-2768):

[0048] MALVLEIFTLLASICWVSANIFEYQVDAQPLRPCELQRETAFLKQADYVPQCAED GSFQTVQCQNDGRSCWCVGANGSEVLGSRQPGRPVACLSFCQLQKQQILLSGYINSTDT SYLPQCQDSGDYAPVQCDVQQVQCWCVDAEGMEVYGTRQLGRPKRCPRSCEIRNRRLLH GVGDKSPPQCSAEGEFMPVQCKFVNTTDMMIFDLVHSYNRFPDAFVTFSSFQRRFPEVS GYCHCADSQGRELAETGLELLLDEIYDTIFAGLDLPSTFTETTLYRILQRRFLAVQSVISGRFRCPTKCEVERFTATSFGHPYVPSCRRNGDYQAVQCQTEGPCWCVDAQGKEMHGTR QQGEPPSCAEGQSCASERQQALSRLYFGTSGYFSQHDLFSSPEKRWASPRVARFATSCP PTIKELFVDSGLLRPMVEGQSQQFSVSENLLKEAIRAIFPSRGLARLALQFTTNPKRLQ QNLFGGKFLVNVGQFNLSGALGTRGTFNFSQFFQQLGLASFLNGGRQEDLAKPLSVGLD SNSSTGTPEAAKKDG...

Embodiment 2

[0053] Example 2: Preparation of TG antibodies

[0054] The TG epitope peptides (1) and (2) obtained in Example 1 were respectively connected with a carrier protein to prepare antigens (1) and (2) for immunization, and the obtained antigens (1) and (2) were used to immunize animals respectively, Thereby, specific monoclonal and polyclonal antibodies are prepared using the antigen (1), and specific monoclonal and polyclonal antibodies are prepared using the antigen (2).

[0055] 1. Preparation of antigens: Tg peptides were respectively connected with carrier protein BSA to prepare TG antigens. Take 5.0 mg of each of the two antigenic peptides described in this application, dissolve them in 1 mL of DMF, and add 5 mg of EDC (dissolved in 50 μL of H) dropwise. 2 O), after stirring for 20 min, the reaction was added dropwise to 5 mg of BSA (dissolved in 1 mL of PBS buffer), 5 mg of NHS was added immediately, and the mixture was stirred at room temperature overnight. The reaction ...

Embodiment 3

[0067] Example 3: Specific identification of human TG antibodies (1) and (2)

[0068] Detected by ELISA. ELISA plates were coated with human Tg protein, GAPDH protein, and neuron-specific enolase NSE as detection antigens, respectively, and the specific reactions of the prepared TG monoclonal antibodies (1) and (2) with different proteins were detected by ELISA. , normal BALB / c mouse serum was used as negative control, and PBS solution was used as blank control.

[0069] Results: Tg monoclonal antibodies (1) and (2) only reacted positively with Tg (P / N > 2.1), respectively, but were negative with GAPDH protein and neuron-specific enolase NSE, indicating that the antibody of the present invention was used. The monoclonal antibodies (1) and (2) prepared from the Tg epitope peptides (1) and (2) have specificity.

[0070] Polyclonal antibodies were identified using the same methods described above for identifying the specificity of monoclonal antibodies.

[0071] The results sh...

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Abstract

The invention discloses a fluorescent immunochromatography test paper for rapidly detecting human thyroglobulin (Tg) to rapidly identify cervical lymph node metastasis of papillary thyroid carcinoma. The test paper detects human Tg by double antibody sandwich method and fluorescent immunochromatography technology. Antibodies are prepared using specific epitope peptides. The invention can quickly and accurately detect the human Tg in the object to be tested, and has the advantages of simple and rapid operation, wide detection range, high specificity and good sensitivity.

Description

technical field [0001] The invention belongs to the field of immune detection, and particularly relates to a human thyroglobulin (Tg) epitope peptide, a Tg-specific antigen prepared with the epitope peptide, and a corresponding monoclonal antibody or polyclonal antibody, and the antibody is used in the preparation of human thyroglobulin (Tg). The use of an in vitro diagnostic kit for papillary thyroid cancer cervical lymph node metastasis, a related in vitro diagnostic kit, and a fluorescent immunochromatographic test strip for rapid intraoperative identification of cervical lymph node metastasis of papillary thyroid cancer and a preparation method thereof. Background technique [0002] Thyroid cancer is the most common malignant tumor among endocrine tumors and head and neck tumors, and its incidence has been the fastest growing solid malignant tumor in the past 20 years. The rapid increase in the incidence of thyroid cancer is mainly attributed to the rapid increase in the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/533G01N33/543G01N33/558
CPCG01N33/533G01N33/54313G01N33/558
Inventor 邹贤周彬李文新施龙顺包建东朱国华高芸王国瑞李秀龙杨克勤
Owner JIANGSU INST OF NUCLEAR MEDICINE
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