Mass spectrum kit for detecting squamous-cell carcinoma antigen feminine cervical carcinoma serum protein and preparation method thereof

A technology for squamous cell carcinoma and cervical cancer, applied in the field of protein detection, can solve the problems of high false negative rate and lack of early monitoring methods

Inactive Publication Date: 2013-06-05
许洋 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the accuracy of the Pap smear is affected by many factors, and the false negative rate is relatively high, about 15% to 40%.
The traditional tumor marker squamous cell carcinoma antigen (SCC-Ag) is of great value in the prognosis assessment of cervical cancer, but it is also found in clinical work that a considerable proportion of cervical cancer patients cannot be detected due to the negative expression of squamous cell carcinoma antigen. For the prognosis assessment of squamous cell carcinoma antigen-negative cervical cancer patients, there is a lack of effective early monitoring methods
However, when we performed protein mass spectrometry analysis, we found that there was no standardized kit for mass spectrometry of clinical squamous cell carcinoma antigen-negative cervical cancer

Method used

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  • Mass spectrum kit for detecting squamous-cell carcinoma antigen feminine cervical carcinoma serum protein and preparation method thereof
  • Mass spectrum kit for detecting squamous-cell carcinoma antigen feminine cervical carcinoma serum protein and preparation method thereof
  • Mass spectrum kit for detecting squamous-cell carcinoma antigen feminine cervical carcinoma serum protein and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1 Distinguishing between normal and squamous cell carcinoma antigen-negative cervical cancer patients and preparation of a mass spectrometry kit

[0069] (1) Experimental method

[0070] The preoperative serum of 77 patients with cervical squamous cell carcinoma (including 44 cases of stage I and 33 cases of stage II, with an average age of 44 years) and 13 patients with CINIII (average age of 41 years). There were 52 healthy subjects, with an average age of 42 years, from a population with normal liver function and renal function tests. Collect 1mL of venous blood from the subject on an empty stomach, immediately after collection, let it stand in a refrigerator at 4°C for 2 hours, centrifuge at 4,000r / min at 4°C for 10 minutes to separate the serum, and centrifuge the serum again at 12,000r / min at 4°C for 5 minutes to remove all residual cell debris and insoluble matter, the serum was divided into 100 μL / tubes on ice, a total of 5 tubes, and stored in a -80°C ...

Embodiment 2

[0095] The double-blind test of embodiment 2 kit

[0096] Screen out several characteristic protein peaks from embodiment 1, 3974,3398, 13761 ± 15Da 3 difference peaks form the test model to SCC-Ag negative expression cervical cancer patient and healthy crowd blind screening test, analyze with this classification method In the mass spectrometry results of 32 cervical cancer samples, 32 of them were correctly differentiated, 0 were wrongly distinguished, and the sensitivity was 100%; 30 of the 32 control samples were correctly distinguished, 2 were wrongly distinguished, and the specificity was 93.75% (see Table 2 ):

[0097] Table 2: Discrimination of test models in biological samples

[0098]

[0099] Note: Sensitivity 100% (32 / 32); Specificity 93.75% (32 / 32)

[0100] The experimental results using C8 and C18 hydrophobic matrix magnetic beads or chips are consistent with the experimental results of the above-mentioned WCX anion matrix magnetic beads or chips.

Embodiment 3CI

[0101] Example 3 CINIII and cervical cancer I, II stage patient polypeptide protein screening

[0102] Comparison of polypeptide proteins between CINIII and patients with cervical cancer stage I and II: After normalization of the protein fingerprints of 13 cervical cancer and 13 CINIII samples, a total of 122 protein peaks were detected in the molecular weight range of 1,500 to 35,000, of which 2 were in There is a significant difference between the cervical cancer group and the control group (P<0.05). The molecular weights, mean values ​​and P values ​​of different peaks are shown in Table 3:

[0103] Table 3: Difference peaks in 13 cases of cervical cancer and 13 cases of cervical CINIII biological samples

[0104] Protein peak Average protein abundance in cervical cancer patients Protein level P value in cervical CINIII patients

[0105] (m / z) x±s mean abundance x±s (P<0.05)

[0106] 9279 15.44±3.97 19.41±4.30 0.0129

[0107] 5944 3.97±1.57 5.28±1.81 0.0257

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Abstract

The invention relates to a magnetic bead supporting substrate method for capturing cervical cancer proteome in biological sample in vitro, which uses magnetic separator to separate magnetic bead and sample, without centrifugation of samples. Then fingerprint technique is used to analyze protein with mass spectrometry method. The invention can be used in kits for testing squamous cell carcinoma negative antigen cervical cancer sample and normal person in vitro and for protein fingerprint or mass spectrometry polypeptide mapping of prognosis. The method is accurate, convenient and fast.

Description

technical field [0001] The invention relates to a new reagent kit for the protein analysis method in the squamous cell carcinoma antigen-negative cervical cancer biological sample. A protein-bound matrix captures biomarkers and detects squamous cell carcinoma antigen-negative cervical cancer biomarkers using quantitatively controlled mass spectrometry. The invention mentioned here relates to the field of protein detection and is a new non-invasive in vitro mass spectrometry detection method. The present invention can be applied to the detection method or kit of the squamous cell carcinoma antigen-negative cervical cancer biomarker combination in the body fluid that has been separated from the human body. Background technique [0002] Both normal function and pathological characteristics of cells depend to some extent on the function of proteins expressed by cells. Therefore, the identification of differences in proteins expressed in the human body can be used for diagnosis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N30/02
Inventor 许洋余传定
Owner 许洋
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