CN104634857A - Application of organic acid molecule as stomach cancer marker - Google Patents
Application of organic acid molecule as stomach cancer marker Download PDFInfo
- Publication number
- CN104634857A CN104634857A CN201310556922.6A CN201310556922A CN104634857A CN 104634857 A CN104634857 A CN 104634857A CN 201310556922 A CN201310556922 A CN 201310556922A CN 104634857 A CN104634857 A CN 104634857A
- Authority
- CN
- China
- Prior art keywords
- stomach
- cancer
- purposes
- acid molecule
- organic acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to the technical field of biological detection, in particular to application of an organic acid molecule as a stomach cancer marker. The invention discloses application of the organic acid molecule as a stomach cancer marker in preparation of stomach cancer detection kits. The organic acid molecule is one of butanoic acid 2-methyl-3-oxo and levulinic acid or a combination thereof. The organic acid molecule provided by the invention has the characteristics of good sensitivity and specificity, simple operation and little time. At the same time, the detection sample adopted by the invention is urine, thus avoiding physical pain brought to patients due to use of tissue and blood samples. Therefore, no matter from patient compliance or from the accuracy of detection results, the organic acid molecule provided by the invention has very good clinical application value.
Description
Technical field
Technical field of biological of the present invention, is specifically related to the purposes of a class organic acid molecule as stomach cancer marker.
Background technology
Cancer of the stomach is as the modal second largest malignant tumour in the whole world, and one of modal malignant tumour of Ye Shi China, accounts for first of the malignant tumor of digestive tract cause of death.The incidence of disease of China's cancer of the stomach is about 20,/10 ten thousand, annual new patients with gastric cancer 400,000 people, death toll 300,000 people, and the ratio that young people gets a cancer of the stomach is in increase, and 15% patient is less than 40 years old young man.Most of patients with gastric cancer is many with non-specific upper gastrointestinal malaise symptoms, therefore carries out to Symptomatic crowd the recall rate that extensive examination could improve cancer of the stomach, improves screening efficiency.Even if but have research prompting Japan its early carcinoma of stomach discovery rate 40%, find when only small part is and utilizes the Large-scale Screening of gastrofiberscope and radiological examination, current most hospital depends on x-ray, and Type B is ultrasonic, endoscopy and living tissue pathological examination etc. are diagnosed tumour.More than check that some is comparatively large to patient trauma, require high to instrument and equipment, be difficult to use in extensive tumor examination or health examination.
And the most frequently used serum tumor marker of the cancer of the stomach being applicable to extensive lesion detection mainly contains CA199, CA724 and CEA etc., the cancer of the stomach in any stage of about 15 ~ 50% can be detected, the verification and measurement ratio for early carcinoma of stomach is then down to about 2 ~ 11%.There is sensitivity and specificity and be difficult to the problems such as unified in these marks, is difficult to become desirable tumor marker and for clinical detection or large-scale crowd examination.This present situation detects to the early molecule of tumour, curative effect and prognostic evaluation, recurrence and the aspect such as transfer monitoring and biological therapy cause certain difficulty.Therefore the new cancer of the stomach detection mark being applicable to Large-scale Screening, Noninvasive detection is needed.
Summary of the invention
The object of the present invention is to provide a kind of cancer of the stomach detection mark being applicable to Large-scale Screening, Noninvasive detection, there is the problem such as sensitivity and specificity disunity to overcome existing clinical detection tumor marker such as the marks such as CA125, CA199, CA724, AFP, CEA.
For reaching this object, the invention discloses a class organic acid molecule as stomach cancer marker purposes in preparation cancer of the stomach detection kit, described organic acid molecule is a kind or their combination in 2-methyl-3-ketobutyric acid (Butanoic acid, 2-methyl-3-oxo), laevulic acid (Levulinic acid).
In some embodiments, described cancer of the stomach detection kit the detection sample that is suitable for be the urine specimen of person under inspection.
In some embodiments, described experimenter is the normal people of hepatic and renal function, causes urine content of target molecules to fluctuate, affect result to reduce because hepatic and renal function is abnormal.
In some embodiments, described urine sample takes from mud-stream urine on an empty stomach in experimenter's early morning, to be conducive to detecting.
In some embodiments, described cancer of the stomach detection kit is applicable to chromatography, mass spectroscopy, chromatograph-mass spectrometer coupling method, wherein chromatography is vapor-phase chromatography (GC), liquid phase chromatography (LC), high performance liquid chromatography (HPLC), chromatograph-mass spectrometer coupling method comprises GC-MS(gas chromatography-mass spectrography) (GC/MS), Liquid Chromatography-Mass Spectrometry (LC/MS), tablets by HPLC-MS (HPLC/MS), gas chromatography tandem mass spectrometry coupling method (GC/MS-MS), Liquid Chromatography-tandem Mass method (LC-MS/MS), high performance liquid chromatography-tandem mass coupling method (HPLC/MS-MS).
On the other hand, the invention also discloses the biomarker of a class for checkout and diagnosis cancer of the stomach, it is characterized in that described biomarker is a kind or their combination in 2-methyl-3-ketobutyric acid (Butanoic acid, 2-methyl-3-oxo), laevulic acid (Levulinicacid).
On the other hand, the invention also discloses a kind of cancer of the stomach detection method, it carries out the quantitative detection of above-mentioned biomarker to the urine specimen of person under inspection by detection method, when the diagnostic test critical value of biomarker content described in experimenter higher than above-mentioned biomarker, experimenter can be diagnosed as patients with gastric cancer.
In some embodiments, when the diagnostic test critical value of above-mentioned biomarker material is respectively: 2-methyl-3 ketobutyric acid 1.159999ug/ml, laevulic acid 0.545333ug/ml.
In some embodiments, described experimenter is the normal people of hepatic and renal function, causes urine content of target molecules to fluctuate, affect result to reduce because hepatic and renal function is abnormal.
In some embodiments, described urine sample takes from mud-stream urine on an empty stomach in early morning, to be conducive to detecting.
In some embodiments, described detection method can select chromatography, mass spectroscopy, chromatograph-mass spectrometer coupling method, wherein chromatography is chromatography is vapor-phase chromatography (GC), liquid phase chromatography (LC), high performance liquid chromatography (HPLC), chromatograph-mass spectrometer coupling method comprises GC-MS(gas chromatography-mass spectrography) (GC/MS), Liquid Chromatography-Mass Spectrometry (LC/MS), tablets by HPLC-MS (HPLC/MS), gas chromatography tandem mass spectrometry coupling method (GC/MS-MS), Liquid Chromatography-tandem Mass method (LC-MS/MS), high performance liquid chromatography-tandem mass coupling method (HPLC/MS-MS), preferred gas chromatography chromatography mass spectrometry.
In some embodiments, described cancer of the stomach is early carcinoma of stomach.
Organic acid molecule of the present invention, as stomach cancer marker, has good sensitivity and specificity, simple to operate, consuming time few, and the present invention simultaneously adopts and detects sample is urine, avoids bringing patient's human body when adopting tissue, blood sample painful.Therefore, be no matter the compliance from patient or the accuracy from testing result, organic acid molecule of the present invention has good clinical value as stomach cancer marker.
Accompanying drawing explanation
Fig. 1. each biomarker hybrid standard product separate colors spectrogram.
Fig. 2. each biomarker molecular mass figure, wherein A:2-methyl-3 oxobutyric; B: laevulic acid.
Fig. 3. each biomarker diagnosis of gastric cancer ROC curve, wherein A:2-methyl-3 oxobutyric; B: laevulic acid.
Fig. 4. biomarker associating ROC curve.
Embodiment
Below in conjunction with specific embodiment, illustrate the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually conveniently condition, or according to the condition that manufacturer advises.Ratio and number percent based on weight, unless stated otherwise.
Embodiment 1: instrument, reagent, preparation of samples and pre-service.
Reagent and instrument: L-2-chlorophenylalanine, ethyl chloroformate, pyridine, absolute ethyl alcohol, NaOH, chloroform, anhydrous sodium sulfate, the above reagent such as 2-methyl-3 ketobutyric acid, laevulic acid is and analyzes pure, produced by Shanghai traditional Chinese medicines chemical reagents corporation.KQ-250DB type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd.'s production).LD5-2A type desk centrifuge (system in Beijing Jing founds hydro-extractor company limited).Agilent company of gas chromatography mass spectrometer TurboMass(U.S. 7890A GC/5975C MS) carry NIST2011 spectrum storehouse and Chemical Software workstation.
Subject sample: random acquisition urine sample 200 parts, wherein 89 parts is Healthy People, and 111 parts is the in-patient (wherein early carcinoma of stomach 33 example) requiring operative treatment because getting a cancer of the stomach.All experimenter's hepatic and renal functions are all normal, and urine sample all takes from mud-stream urine on an empty stomach in early morning.Cancer of the stomach experimenter all confirms through electronic gastroscopy or excision pathologic finding.
Sample pretreatment: after sample is risen to room temperature by low temperature refrigerator taking-up nature, carry out 10000rpmX20min, centrifugal.Draw Supernatant samples 300ul, adding distil water 300ul, internal standard compound (0.1mg/mL L-2-chlorophenylalanine) 100ul, absolute ethyl alcohol 40ul, pyridine 100ul, add lid and carry out esterifying carboxyl group reaction.Add ethyl chloroformate (ECF) 50ul, add a cover vibration 10S, carry out amino reaction.Ultrasonically promoted reaction 1min, adds chloroform 300ul and extracts.3000rpm X5min after vibration 30S is centrifugal.Add the NaOH of 100ul7mol/L, regulate pH=9 ~ 10, then add ethyl chloroformate (ECF) 50ul, add a cover, vibrate 10S, ultrasonically promoted reaction 1min, carry out secondary reaction.After vibration 30S, 3000rpm × 10min is centrifugal.Suck upper aqueous layer, chloroform layer adds a small amount of anhydrous sodium sulfate for dehydration, and be available on the machine detection.
Embodiment 2: instrument scan method sets
Scan mode is that full ion scan is in conjunction with Selective ion mode (SIM ion) monitoring scanning.SIM ion is set as 2-methyl-3 ketobutyric acid 74m/z, laevulic acid 99m/z, and sample is first in the upper separation of gas-phase chromatographic capillary column (Agilent J & W DB-5MS, 30m × 0.25mm × 0.25 μm).And adopting programmed temperature gas chromatography technology, initial temperature is 80 DEG C, rises to 140 DEG C with 10 DEG C/min speed, then rises to 280 DEG C with 4 DEG C/min speed, and keeps 2min.Increase column temperature gradually in detachment process, make all components can under respective optimum temperature wash-out.Mass spectrometer electronics bombarding energy is 70eV, and ion source temperature is 200 DEG C, and injector temperature is 260 DEG C, and carrier gas is helium, and column flow rate is 1ml/min.All data are collected under mass-to-charge ratio 30-550m/z.Each sample introduction scanning residence time is 100ms, and the solvent delay time is 3.5min.In the sample detected, substance migration standard substance mass spectrogram is qualitative.In sample, the concentration of material uses quantified by external standard method.
Embodiment 3: the foundation of standard quality spectrogram and typical curve.
Preparation biomarker standard items mixed solution, be diluted to 10 concentration gradients successively, according to the pre-service of embodiment 1 method, embodiment 2 method sample introduction, result is as table 1.Each biomarker typical curve is linearly good, in this, as quantitative basis.Each biomarker retention time is shown in Fig. 1, and (sequence number is respectively 1:2-methyl-3 oxobutyric; 2: laevulic acid), corresponding mass spectrogram is shown in 2, in this, as qualitative foundation.
Table 1 each target molecule retention time (RT), the typical curve range of linearity (Linear range) and the right (r of Linear Quasi
2)
Embodiment 4: person under inspection's urine detection
200 parts of urine samples are processed according to the method for embodiment 1, according to embodiment 2 method sample introduction, obtains the chromatogram of each sample.The mass spectrogram obtained according to embodiment 3 subsequently carries out qualitative, and typical curve carries out quantitatively, and data processing platform (DPP) is that instrument carries Chemical Software workstation.After comprehensive each sample target compound content, statistical study is carried out with spss19, in its results sample there is significant difference in 2-methyl-3 ketobutyric acid, laevulic acid Small molecular in cancer of the stomach group and normal healthy controls group, its mean value and standard deviation are in table 2, and in early days in cancer of the stomach group and normal healthy controls group, these micromolecular compound content also significant difference (table 3).This shows that in urine, 2 kinds of small molecule biomarkers thing content not only as the biomarker of cancer of the stomach, can also can be played an important role in early gastric caacer early warning.
Biomarker detected value statistics (ug/ml) in table 2. cancer of the stomach group and normal healthy controls group urine
Table 3. early carcinoma of stomach and normal healthy controls group urine two kinds of biomarker detected values are added up (ug/ml)
Utilize ROC tracing analysis and area under curve (AUC) algorithm to obtain 2 kinds of single variablees of biomarker in sample respectively and differentiating that the ROC curve of cancer of the stomach and non-cancer is shown in Fig. 3, corresponding ROC area under curve, youden index, diagnosis dividing value are in table 4.
Table 4
Utilize SPSS19 to 2 Index Establishment Logistic regression models, its Logistic regression forecasting equation is p=1/ [1+e
-+0.970X1+0.717X2-1.249)], p is the prediction probability obtained according to each person under inspection 2 index calculate.Calculate all person under inspection's prediction probability values, and set up Joint Index ROC curve (Fig. 4) with this value, ROC area under curve is 0.718, determine that the sensitivity that its best dividing value point is corresponding is 0.757 according to Youden index maximal value (0.338), specificity 0.581, its corresponding diagnosis dividing value be p=0.4958702. namely as person under inspection's prediction probability value P>0.49582, think that this person under inspection suffers from cancer of the stomach, its sensitivity is 0.757, specificity 0.581.
The above results shows the method for the invention, by detecting 2 kinds of key small molecules of urine, not only having the effect differentiating cancer of the stomach and non-cancer, can also distinguish early carcinoma of stomach and health volunteer, have good sensitivity and specificity.
Scope of the present invention is not by the restriction of described specific embodiments, and described embodiment only for as the single example of illustrating various aspects of the present invention, also comprises method and the component of functional equivalent in the scope of the invention.In fact, except content as herein described, those skilled in the art can easily grasp multiple improvement of the present invention with reference to description above and accompanying drawing.Described improvement also falls within the scope of appended claims.Every section of list of references mentioned above is listed in herein as a reference all in full.
Claims (9)
1. a class organic acid molecule is as the purposes of stomach cancer marker in preparation cancer of the stomach detection kit, and described organic acid molecule is one of in 2-methyl-3-ketobutyric acid, laevulic acid or their combination.
2. purposes as claimed in claim 1, is characterized in that organic acid molecule is 2-methyl-3-ketobutyric acid, laevulic acid.
3. purposes as claimed in claim 1, it is characterized in that described cancer of the stomach detection kit the detection sample that is suitable for be the urine specimen of person under inspection.
4. purposes as claimed in claim 3, is characterized in that described experimenter is the normal people of hepatic and renal function.
5. purposes as claimed in claim 3, is characterized in that described urine sample takes from mud-stream urine on an empty stomach in experimenter's early morning.
6. purposes as claimed in claim 1, is characterized in that described cancer of the stomach detection kit is applicable to chromatography, mass spectroscopy, chromatograph-mass spectrometer coupling method.
7. purposes as claimed in claim 1, is characterized in that described chromatography is vapor-phase chromatography (GC), liquid phase chromatography (LC), high performance liquid chromatography (HPLC).
8. purposes as claimed in claim 1, is characterized in that described chromatograph-mass spectrometer coupling method is GC-MS(gas chromatography-mass spectrography) (GC/MS), Liquid Chromatography-Mass Spectrometry (LC/MS), tablets by HPLC-MS (HPLC/MS), gas chromatography tandem mass spectrometry coupling method (GC/MS-MS), Liquid Chromatography-tandem Mass method (LC-MS/MS), high performance liquid chromatography-tandem mass coupling method (HPLC/MS-MS).
9. a class is used for the biomarker of checkout and diagnosis cancer of the stomach, it is characterized in that described biomarker is a kind or their combination in 2-methyl-3-ketobutyric acid, laevulic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310556922.6A CN104634857A (en) | 2013-11-11 | 2013-11-11 | Application of organic acid molecule as stomach cancer marker |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310556922.6A CN104634857A (en) | 2013-11-11 | 2013-11-11 | Application of organic acid molecule as stomach cancer marker |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104634857A true CN104634857A (en) | 2015-05-20 |
Family
ID=53213844
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310556922.6A Pending CN104634857A (en) | 2013-11-11 | 2013-11-11 | Application of organic acid molecule as stomach cancer marker |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104634857A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102782157A (en) * | 2010-04-30 | 2012-11-14 | 香港中文大学 | Marker for gastric cancer and method for detecting gastric cancer |
| CN103224922A (en) * | 2013-04-11 | 2013-07-31 | 西安交通大学 | New stomach cancer marker, detection method and applications thereof |
-
2013
- 2013-11-11 CN CN201310556922.6A patent/CN104634857A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102782157A (en) * | 2010-04-30 | 2012-11-14 | 香港中文大学 | Marker for gastric cancer and method for detecting gastric cancer |
| CN103224922A (en) * | 2013-04-11 | 2013-07-31 | 西安交通大学 | New stomach cancer marker, detection method and applications thereof |
Non-Patent Citations (1)
| Title |
|---|
| KIMBERLY KAPLAN ET.AL: "Metabolic differences among melanoma and two prostate cancer cell lines by electrospray ion mobility mass spectrometry", 《INTERNATIONAL JOURNAL FOR ION MOBILITY SPECTROMETRY》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lv et al. | Identification of possible biomarkers for breast cancer from free fatty acid profiles determined by GC–MS and multivariate statistical analysis | |
| Wang et al. | NMR-based metabolomic techniques identify potential urinary biomarkers for early colorectal cancer detection | |
| Fan et al. | Identification of metabolic biomarkers to diagnose epithelial ovarian cancer using a UPLC/QTOF/MS platform | |
| CN108414660B (en) | Application of group of plasma metabolism small molecule markers related to early diagnosis of lung cancer | |
| Xiang et al. | Volatile organic compounds in human exhaled breath to diagnose gastrointestinal cancer: a meta-analysis | |
| Zhang et al. | Serum unsaturated free fatty acids: a potential biomarker panel for early-stage detection of colorectal cancer | |
| CN104634907A (en) | Application of amino acid molecular combination as stomach cancer marker | |
| CN105044361A (en) | Diagnosis marker suitable for early-stage esophageal squamous cell cancer diagnosis and screening method of diagnosis marker | |
| CA2772688A1 (en) | Methods for the diagnosis of colorectal cancer and ovarian cancer health states | |
| CN112162050B (en) | Application of MIT and/or DIT as thyroid cancer marker and kit | |
| CN106370744A (en) | Application of 8-hydroxydeoxyguanosine as urine marker | |
| Tyan et al. | Urinary protein profiling by liquid chromatography/tandem mass spectrometry: ADAM28 is overexpressed in bladder transitional cell carcinoma | |
| US20200173996A1 (en) | Device for diagnosing colorectal cancer and method for providing colorectal cancer diagnosis information | |
| CN101382537A (en) | Stomach cancer diagnostic method | |
| Kałużna-Czaplińska et al. | Current applications of chromatographic methods for diagnosis and identification of potential biomarkers in cancer | |
| Li et al. | A pilot study for colorectal carcinoma screening by instant metabolomic profiles using conductive polymer spray ionization mass spectrometry | |
| CN112986441A (en) | Tumor marker screened from tissue metabolism contour, application thereof and auxiliary diagnosis method | |
| CN105116000B (en) | Nuclear magnetic resonance model and preparation method for detecting stomach cancer associated metabolic small molecule | |
| Oxner et al. | The versatility and diagnostic potential of VOC profiling for noninfectious diseases | |
| JP2013246080A (en) | Colorectal cancer inspection method | |
| CN109811033A (en) | ACOX1 is preparing the application in ICP auxiliary diagnostic box as detection target spot | |
| CN115420839A (en) | Application of marker in prediction of prostate cancer and related product | |
| CN110749732B (en) | Blood metabolite marker for diagnosing multiple myeloma and application thereof | |
| CN104634857A (en) | Application of organic acid molecule as stomach cancer marker | |
| CN109061179A (en) | Application of the amino acid combined factor in building colorectal cancer hematology diagnostic model |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150520 |
|
| RJ01 | Rejection of invention patent application after publication |