Ovarian-cancer marker detection kit based on PRM (Parallel Reaction Monitoring) detection and detection method

A detection kit and detection method technology, applied in the field of ovarian cancer marker detection, can solve the problems of time-consuming, poor sensitivity and specificity, complicated operation, etc., and achieve the effect of high safety, strong specificity, and simple operation

Inactive Publication Date: 2018-06-12
上海中科新生命生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the operation of the OVA1 test kit is complicated and time-consuming, and it

Method used

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  • Ovarian-cancer marker detection kit based on PRM (Parallel Reaction Monitoring) detection and detection method
  • Ovarian-cancer marker detection kit based on PRM (Parallel Reaction Monitoring) detection and detection method
  • Ovarian-cancer marker detection kit based on PRM (Parallel Reaction Monitoring) detection and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment one: chromatographic analysis column.

[0032] Such as figure 1 A chromatographic analysis column suitable for the detection of ovarian cancer PRM is shown, which includes a column tube (1), a filler (2) and a tip (3), the filler (2) is filled in the column tube (1), and the tip (3) Cover one end of the column tube (1); the length of the column tube (1) is 120mm, the outer diameter is 360um, and the inner diameter is 75um; the filler (2) is a C18 reverse phase filler with a particle size of 3um .

Embodiment 2

[0033] Example 2: Ovarian Cancer Marker Detection Kit

[0034] A detection kit for ovarian cancer markers based on PRM detection, including reagents for conventional mass spectrometry detection, heavy isotope-labeled standard peptide test tubes, the above-mentioned chromatographic analysis column, and an aqueous solution containing 0.1% formic acid; the heavy isotope-labeled standard peptide The test tube contains ten unique sequences among thyroxine, apolipoprotein A–1, β2-microglobulin (Beta2M), transferrin, and cancer antigen 125, where each protein has two peptide sequences, namely VEHSDLFSK, For VNHVTLSQPK, EFQLFSSPHGK, HQTVPQNTGGK, GSPAINVAVHVFR, AADDTWEPFASGK, DLATVYVDVLK, DYVSQFEGSALGK, LGIWDDFIPK, and ELPLLSPPQDK, the heavy isotope-labeled amino acids were located on arginine and lysine at the C-terminal of the peptide sequence, respectively.

[0035] As a preferred embodiment, the concentration of each standard peptide in the heavy isotope-labeled standard peptide te...

Embodiment 3

[0036] Example 3, detection of ovarian cancer markers.

[0037] Step S10, take a heavy isotope-labeled standard peptide test tube, add 90-110uL of dissolving buffer into it, shake and mix, use a centrifuge to centrifuge at a speed of 13000g for 60s, and take the supernatant for later use;

[0038] Step S20, taking 0.1 mg of serum proteolytic peptides and adding 100 uL of dissolving buffer, shaking and mixing to make a serum peptide solution for later use;

[0039] Step S30, taking 2uL of the serum peptide solution prepared in step S20, and adding 10uL of the heavy isotope-labeled standard peptide solution prepared in step S10, mixing evenly, taking 4uL for chromatographic injection;

[0040] The chromatographic separation gradient was set as follows:

[0041] time (min)

[0042] The PRM detection ion table is set as follows:

[0043] M / Z

[0044] The mass spectrometer was set up as follows:

[0045]First-level mass spectrometry, scanning range: 500-800m / ...

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Abstract

The invention provides an ovarian-cancer marker detection kit based on PRM (Parallel Reaction Monitoring) detection and a detection method thereof and belongs to the technical field of ovarian-cancermarker detection. The ovarian-cancer marker detection kit is used for rapidly and accurately detecting the contents of 5 types of markers and comprises a reagent for conventional mass-spectrometric detection, a heavy-isotope-labeled standard peptide fragment test tube and a special chromatographic analysis column; 10 artificially-synthesized heavy-labeled isotope peptide fragments of 5 ovarian-cancer protein markers are doped into serum, and a PRM technology is used for carrying out absolute quantitative analysis on the 5 ovarian-cancer protein markers which are endogenous in the serum so as to achieve the purpose of helping diagnosis of the ovarian cancer. The ovarian-cancer marker detection kit provided by the invention has the advantages of simple operation, short consumed time, short experimental period, strong specificity and high sensitivity.

Description

technical field [0001] The invention belongs to the technical field of detection of ovarian cancer markers, and in particular relates to a detection kit for ovarian cancer markers based on PRM detection, and also relates to a detection method using the detection kit. Background technique [0002] Malignant tumor of the ovary is one of the common malignant tumors of the female reproductive organs. Its incidence rate ranks third after cervical cancer and uterine body cancer, accounting for about 4% of female malignant tumors in the whole body. However, the number of deaths due to ovarian cancer ranks first in all types of gynecological tumors, posing a serious threat to women's lives. [0003] Parallel reaction monitoring (Parallel reaction monitoring, PRM) Parallel reaction monitoring belongs to targeted MS / MS analysis, and will continuously record all fragment ion spectra of each target precursor ion during the entire liquid phase separation process. Compared with tradition...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/60G01N30/88
CPCG01N30/06G01N30/6073G01N30/88
Inventor 陈薇阮宏强
Owner 上海中科新生命生物科技有限公司
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