CN102565267B - Kit for screening colorectal cancer - Google Patents
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Abstract
The invention discloses a kit for screening colorectal cancer, which comprises a device and a comparison card, wherein the device is used for detecting the content of phosphatide; the comparison card is established by utilizing the content of phosphatide; and the phosphatide is SPC (sphingosylphosphorylcholine), 14:0 LPC (lysophosphatidylcholine), 16:0 LPC, 18:2 LPC, 18:1 LPC, 18:0 LPC, 20:4 LPC, 20:0 LPC, 22:6 LPC and 22:0 LPC. A test of the kit proves that an effective early diagnosis molecule mark for the colorectal cancer is developed, a molecule detection model is established, the detection sensitivity and specificity can reach 88% and 80% according to the model, and a distinguishing effect for comparing the colorectal cancer with the normal state obtained by utilizing the lipid molecules is better than the one obtained by using the marks for researching the early diagnosis of the colorectal cancer in recent years.
Description
Technical field
The present invention relates to biological technical field, relate in particular to a kind of kit of examination colorectal cancer.
Background technology
Colorectal cancer rises at global incidence of disease straight line in recent years, exceedes every year in the last few years million people and dies from colorectal cancer.Due to generation and the habits and customs of colorectal cancer, particularly eating habit is closely related, and therefore, in recent years along with the raising gradually of China's living standards of the people, the incidence of disease and the fatal rate of colorectal cancer rapidly increase in China.Research shows, the early diagnosis of colorectal cancer is particularly crucial, and the five-year survival rate of early detection colorectal cancer patients is up to 90%, and colorectal cancer early detection rate less than 40% at present, once shift colorectal cancer patients survival rate less than 10%." goldstandard " of current diagnosis colorectal cancer is Sigmoidoscope, generally acknowledge at present, the age be greater than 50 years old should accept per capita the examination of intestines mirror.The authoritative departments such as the U.S. and Europe point out, for there is familial medical history occurred frequently or due to the high people of the ill risk of personal habits the examination of intestines mirror particularly important.But China does not also possess the condition of extensively carrying out the survey of intestines microscopy in health check-up at present, experimenter's misery is also self-evident.In China's health check-up, extensively carrying out at present Sigmoidoscope detects unrealistic.Therefore, in screening blood plasma, colorectal cancer early diagnosis molecular labeling becomes one of study hotspot now.
People have determined and have been present in a large number the biomarker in colon, ight soil or serum, as carcinomebryonic antigen (CEA) etc., come early diagnosis and prediction for colon cancer, but the sensitivity of the screening method that most is recommended and specificity are all below 50%.Recently, also in succession reported the colorectal cancer molecular labeling in some tissues, in ight soil and in serum, but its effect is also far below Sigmoidoscope.For example, if served as a mark with the special site that methylates of faeces DNA, in the time that specificity reaches 90%, sensitivity only has 50%.And this detection easy to detect not as blood marker certainly, therefore at present more medical personnel wishes to develop the mark of diagnosis of colorectal carcinoma in blood.Studies have reported that, serum colon cancer specific antigen CCSA-3 and CCSA-4 may become the mark of diagnosis of colorectal carcinoma, and it detects effect and also needs to confirm.Annual approximately report has thousands of kinds of tumor molecular markers, but being finally applied to clinical molecular marker does not almost have, the defect that their exist concentrates on sample size less (< 30), not in not agnate or independent experiment by certain etc.More crucial, be difficult to be promoted by these molecular markers at the disunity of sample collection, pre-service and target molecule extractive process Plays.
To sum up, because intestines mirror expense is higher, the degree of reason reduces patient would like to do this inspection such as painful large and operation requirements is high, causes a lot of patients to be dragged to and become incurable disease late period.Therefore need to develop in a hurry a kind of can be applicable to colorectal cancer early screening simple and reliable, without wound or the large flux inspection method of Wicresoft.
There is bioactive lysophosphatide molecule and day by day cause scientist's concern in tumour generation and developing effect.This some of them lysophosphatide molecule, such as LPC, sphingosylphosphocholine (sphingosylphosphorylcholine, SPC), lysophosphatidic acid (lysophosphatidic acid, LPA), sphingosine-1-phosphate (sphingosine-1-phosphate, S1P) and sphingomyelins (sphingomyelin, SM) can regulate such as cell proliferation, migration, form occur and be adsorbed in various activities.Therefore these lysophosphatide molecules can participate in Angiogenesis, wound healing, immunity, atherosclerotic generation and the adjusting that becomes the processes such as knurl.The metabolism of these lipid molecules and above-mentioned physiological and pathological process are closely bound up, and wherein SPC and SM all can be converted into S1P, and LPC can generate LPA by ATX catalysis, and S1P and LPA are very remarkable to the regulatory function of tumour as functional activity lipid molecule.Along with biologically active lipid molecule and cancer-related understanding deeply and the progress of lipid molecule detection means, some biologically active lipid molecules are considered to serve as the biomarker of early diagnosis of tumor.Based on lipid molecule, particularly important physiology and the pathology sense of biologically active lipid molecule, has proposed the concept of lipidomics and function lipidomics in the world recently.Except existing genome and proteomics, the research and development of lipidomics will play an important role in the early diagnosis and therapy of tumour.In addition, tumour occur the metabolism of early stage biologically active lipid molecule extremely can be from people's body fluid (blood serum/blood plasma) direct-detection to.By comparison, traditional tissue samples detects that not only wound is large, and tumour is difficult to accomplish early diagnosis.Compare with protein marker with gene, as metabolic marker thing, the detection of biologically active lipid molecule is more convenient, quick, is suitable for clinical expansion.Therefore, set up sensitive, accurate and efficient biologically active lipid analysis method, screening, for the TS biologically active lipid molecule of difference label, has great importance to early diagnosis of tumor.
Be different from proteins and peptides, exploitation is directed to simple lysophosphatide molecule, as very difficult in the antibody of LPA and LPC.Therefore simple ELISA detection method is difficult to be suitable for for these lysophosphatide molecules.Some traditional lipid analysis methods were once successfully applied to the analyzing and testing of lysophosphatide, as thin-layer chromatography (TLC), phosphorous determination, and gas chromatography (GC) etc.But, the analytic process complexity having in these methods, workload is large, the poor sensitivity having, the low lysophosphatide of content in analysing body fluid efficiently, and without any a kind of method can be simply for separating of with the hypotype of analyzing lysophosphatide.
Summary of the invention
The object of this invention is to provide the kit of a kind of examination or auxiliary examination colorectal cancer.
The kit of examination provided by the invention or auxiliary examination colorectal cancer, comprises that the comparison of setting up for detection of the device of content of phospholipid with by described content of phospholipid blocks;
Described phosphatide is SPC, 14:0LPC, 16:0LPC, 18:2LPC, 18:1LPC, 18:0LPC, 20:4LPC, 20:0LPC, 22:6LPC and 22:0LPC.
The described comparison card of being set up by content of phospholipid is following 1) or 2):
1) be that comparison card A and comparison block any one in B;
2) comparison card A and comparison card B;
Described comparison card A is made up of straight line 1, horizontal ordinate 1 and ordinate 1;
Described straight line 1 is 0.023 × saturated LPC total amount+5.96 × 18:2LPC%+17.06 × SPC total amount=9.66;
Described horizontal ordinate 1 is 0.023 × saturated LPC total amount+5.96 × 18:2LPC%;
Described ordinate 1 is SPC total amount (content);
Above-mentioned straight line 1 is the linear function of ordinate 1 about horizontal ordinate 1;
Described comparison card B is made up of straight line 2, horizontal ordinate 2 and ordinate 2;
Described straight line 2 is 11.41 × 18:2LPC%+7.43 × 18:1LPC%+0.024 × saturated LPC total amount=14.58;
Described horizontal ordinate 2 is 11.406 × 18:2LPC%+7.43 × 18:1LPC%;
Described ordinate 2 is saturated LPC total amount;
Above-mentioned straight line 2 is the linear function of ordinate 2 about horizontal ordinate 2;
Described 18:2LPC% is the number percent that 18:2LPC content accounts for unsaturated LPC total amount;
Described 18:1LPC% is the number percent that 18:1LPC content accounts for unsaturated LPC total amount.
Described saturated LPC total amount is the summation of 14:0LPC, 16:0LPC, 18:0LPC, 20:0LPC and 22:0LPC content;
Described unsaturated LPC total amount is the summation of 18:2LPC, 18:1LPC, 20:4LPC and 22:6LPC content;
The unit of above-mentioned each content of phospholipid is micro M.
The described device for detection of content of phospholipid is LC-MS instrument.
Described LC-MS instrument is Applied Bio systems S ciex 3200Qtrap
tMmass spectrometer, Applied Biosystems Sciex, Ontario, Canada, it detects mobile phase used is that volume ratio is methyl alcohol, water, the ammonia water mixture of 90: 10: 0.1;
The flow velocity of the mobile phase that described detection adopts is 0.2ml/min,
While detecting LPC, SPC, L-PAF and the each subclass of SM, do not need pillar, the pillar model that detects LPA, S1P is: C18HPLC column (TARGA C18 5 μ M, 2.1mm ID × 10mm TR-0121-C185, Higgins Analytical, Southborough, MA).
Described content of phospholipid is content of phospholipid in vitro blood.
Described kit is made up of the described device for detection of content of phospholipid and the described comparison card of being set up by described content of phospholipid.
The application of described kit in the product of preparation detection sample to be tested colorectal cancer is also the scope of protection of the invention.
Described sample to be tested derives from people, specifically derives from Asian, is especially preferably Chinese.
Of the present invention experimental results show that, the present invention's (each 120 examples of normal control and tumor patient) in Chinese population has furtherd investigate the content (LPC of a series of phospholipid molecules, LPA, SPC, S1P, SM and haemolysis platelet factor L-PAF), particularly contain the lipid molecule (LPC of choline group, SM, and SPC) content of various subclass in blood plasma, deeply probe into the variation of these lipids in colorectal cancer, and then develop effective colorectal cancer early diagnosis molecular labeling, set up Molecular Detection model, the sensitivity and the specificity that detect according to this model can reach 88% and 80% (or 83% and 86%), utilize these lipid molecules the differentiation effect of colorectal cancer and normal control to be better than much studying in the last few years the mark of colorectal cancer early diagnosis.
In screening of the present invention, be basis with Electro-spray ionization mass spectrometry (ESI-MS, triple level Four tandem mass spectrometers), developed the technology of lysophosphatide molecule in quantitative detection human plasma.The advantage of Mass Spectrometer Method is: (1) is highly sensitive, reproducible; (2) it not only can detect a large amount of different types of lipid molecules simultaneously, and can differentiate the different subtype of same lipid molecule; (3) detect plasma sample and there is the advantages such as sampling is convenient, wound is little, and required sample size only has 10-50 μ l; (4) utilize a self-actuated sampler, this detection technique just can become very robotization.Therefore, this detection method has very large potentiality in clinical practice.Application correlation technique, 2007, first the Xu Yan of indiana ,US university professor reported that LPC can be used as the colorectal cancer early diagnosis marker of American population in laboratory.But for early diagnosis of cancer molecular labeling, most important across race and crowd's independent detection, there is very big difference in most molecular labelings in population.
Result of study of the present invention shows that a series of phospholipid molecules that contain choline group can be used as the molecular labeling of examination colorectal cancer.The metabolism of these lipid molecules plays a key effect for the process of colorectal cancer.Have and exceed 100 every year, 000 section of article about " cancer marker " is delivered, but these molecular markers are finally applied to the few of clinical examination, there is many difference to the testing result of same mark in different experiments chamber, the detection method of the difference of sample collection mode difference, extracting mode and complexity etc. is all the reason that these marks are difficult to be employed.And of the present invention to these containing the lipid molecule testing process of choline group, sample, extract and detect highly standardized, and utilize tens microlitre blood plasma and the simple organic reagent just can the content of these lipid molecules of accurate quantitative analysis in blood plasma, can carry out in routine physical examination, and set it as the screening indexes of colorectal cancer early diagnosis, refer to that blood chemical examination is just likely for diagnosis of colorectal carcinoma provides preliminary foundation.
Brief description of the drawings
Fig. 1 is saturated LPC, unsaturated LPC, total LPC and (16:0+18:0) the content trend of SM content in normal control and colorectal carcinoma patient body
Fig. 2 is for utilizing LPC to distinguish normal control and colorectal cancer patients model in Chinese population
Fig. 3 is for utilizing LPC and SPC to distinguish normal control and colorectal cancer patients model in Chinese population
Embodiment
The experimental technique using in following embodiment if no special instructions, is conventional method.
Material, reagent etc. used in following embodiment, if no special instructions, all can obtain from commercial channels.
The structure of the kit of embodiment 1, examination colorectal cancer
One, the foundation of molecular model
1, blood collection, lipid molecule extract and detect
1) research object
All sample collections are in May, 2007-2010 year May, and sample is from hospital general of the Second Artillery Force of the entire PLA, Beijing Hospital, the second affiliated hospital of Peking University.Get patient blood on an empty stomach morning and carry out pre-service.This project is through Institutional Review Board examination & approval, all subjects signed Informed Consent Forms.
Table 1 has been summarized the basic condition of research object: in tested healthy sample and tumor sample, masculinity proportion is respectively 68% and 59%; The age of normal healthy controls and tumor patient is respectively 55.2 ± 7.5 and 55.7 ± 11.8, and the sampling age does not have significant difference.
Table 2 has been summarized the basic condition of the 120 routine tumor samples of getting: 53% (64/120) is colon tumor, and all the other samples are mostly rectal neoplasm.Exceeding 75% tumour is T3/T4 phase (93/120), and only about half of tumour is N0 (58/120,48%).From differentiation degree, most tumours are middle height differentiation (82/120,69%).
Basic condition table 2 cancer patient's of table 1 research object basic condition
2) blood collection, lipid molecule extract and detect
In the pipe that contains EDTA, collect above-mentioned 1) the in vitro whole blood of sample, room temperature (25 DEG C) 1, centrifugal 15 minutes of 750g, is distributed into Blood plasma in vitro on the pipe of silicidation ,-80 degree Refrigerator stores, avoid multigelation.
The extraction (lipid extraction method 1) of LPC, SPC, SM and L-PAF: add 0.5ml PBS (Hyclone company in glass centrifuge tube, SH30256.01B), add 100 μ l 12:0LPC (avanti polar lipids 855475P, 100mM), add 3ml methyl alcohol/chloroform (volume ratio is 2: 1).In above-mentioned solution, add 10 μ l Blood plasma in vitro obtained above, after whirlpool mixes, place on ice 10 minutes.Add 1ml chloroform, 1.3ml distilled water, mixes, (25 DEG C) 1 are mixed in chamber, and centrifugal 10 minutes of 750g, takes off a layer organic phase, nitrogen dries up, and dissolves by 1ml methanol/water (9: 1), gets machine testing LPC, SPC, SM and L-PAF and each subclass content thereof on 200 μ l.
The extraction (lipid extraction method 2) of LPA and S1P: add 0.5ml PBS in glass centrifuge tube, add 100 μ l 14:0LPA (avanti polar lipids 857120X, 5 μ M), add 3ml methyl alcohol/chloroform (2: 1).In above-mentioned solution, add 50 μ l blood plasma, and 15 μ l 6N HCl, after mixing, places on ice 10 minutes whirlpool.Add 1ml chloroform, 1.3ml distilled water, mixes, room temperature 1, and centrifugal 10 minutes of 750g, takes off a layer organic phase, and nitrogen dries up, and dissolves by 150 μ l methanol/water (9: 1), gets machine testing LPA and S1P and each subclass content thereof on 150 μ l.
Above-mentioned upper machine testing all adopts LC-MS system: LC-MS instrument Applied Biosystems Sciex 3200QtrapTM massspectrometer, Applied Biosystems Sciex, Ontario, Cahada, mobile phase is: methanol/water/ammoniacal liquor (90: 10: 0.1); The flow velocity of described detection content of phospholipid mobile phase is 0.2ml/min, measure LPC, SPC, L-PAF and the each subclass of SM and do not need pillar, the pillar model of measuring LPA and the each subclass of S1P is: C18HPLC column (TARGA C18 5 μ M, 2.1mm ID × 10mm TR-0121-C185, Higgins Analytical, Southborough, MA).
The standard items of series phosphatide are all purchased from Avanti Polar Lipids company (in table 3), and the catalog number of portioned product is as shown in table 3 below:
The standard items of table 3 lipid
| Name | Catal0gue N0. |
| 12:0LPC | 855475P |
| 14:0LPC | 855575P |
| 16:0LPC | 855675P |
| 18:2LPC | 790648P |
| 18:1LPC | 845875C |
| 18:0LPC | 855775C |
| 20:0LPC | 855777C |
| 20:4LPC | Synthesized by the said firm |
| 22:0LPC | 855779C |
| 22:6LPC | Synthesized by the said firm |
| SPC | 860600P |
| lyso-PAF | 878119C |
| 14:0LPA | 857120X |
| 16:0LPA | 857123P |
| 18:1LPA | 857130C |
| 18:0LPA | 857128X |
| S1P | 860492P |
| 16:0SM | 860061C |
| 18:0SM | 860062C |
2, experimental result
1) detected the impact of age on content of phospholipid in the blood plasma detecting
Generally acknowledge at present, the morbidity of colorectal cancer is directly related with the age, and the age is larger, and incidence probability is higher, if but with the age as colorectal cancer early diagnosis label, sensitivity has 79.8%, but specificity only has 23.9%.And all lipid variation tendencies that the present invention detects and age, therefore these lipid molecules that contain choline group were diagnostic markers separate with the age without direct relation.
2) relation between content and the sex of phospholipid molecule in blood plasma
Testing result is as described in Table 4, can find out, if carry out t inspection, using p=0.01 as standard, male sex 18:2LPC, 18:1LPC and the total amount of unsaturated LPC in blood plasma are a little more than women, and difference is also not obvious, and therefore the content of these phospholipid molecules and sex are also without obvious relation.The unit of following each content of phospholipid is micro M.
The gender differences of the different lipid molecules of table 4
p-value from the Wilcoxon rank sum test
3) content of phospholipid molecule (unit is micro M) in blood plasma.
Detect the content of following phospholipid molecule and each subclass in whole samples: SPC, lyso-PAF, 14:0LPC, 16:0LPC, 18:2LPC, 18:1LPC, 18:0LPC, 20:4LPC, 20:0LPC, 22:6LPC, 22:0LPC, 16:0SM and 18:0SM.
By normal healthy controls group, compared with cancer group, SPC, lyso-PAF, 14:0LPC, 16:0LPC, 18:2LPC, 18:1LPC, 18:0LPC, 20:4LPC, 20:0LPC, 22:6LPC and the 22:0LPC content in cancer patient blood plasma is all significantly lower than Normal group (table 5).Saturated LPC, unsaturated LPC and total LPC content all significantly decline in cancer patient blood plasma.This wherein, the downtrending of 16:0LPC, 18:2LPC, 18:1LPC, 18:0LPC, 20:0LPC is consistent with the downtrending of American population.Thereby show that the content of these phospholipid molecules in blood plasma likely becomes molecular labeling shared in not agnate or different crowd.And 16:0SM and the 18:0SM content in normal control and tumor patient blood plasma does not have significant difference (Fig. 1).The content of all phospholipid molecules detecting and tumour by stages with location-independent (data now shown).Detect the difference in group based on these phospholipid molecules in difference, will set up the molecular model of distinguishing normal VS tumour (colorectal cancer).
The content of the different lipid molecules of table 5 in normal control and colorectal carcinoma patient
p-value from the Wilcoxon rank sum test
3, set up molecular model
According to the above results, set up following molecular model, be used for distinguishing China's Healthy contrast and colorectal cancer patients:
The formula of model 1: [0.023 × saturated LPC total amount+5.96 × 18:2LPC%+17.06 × SPC total amount=9.66];
The formula of model 2: [11.41 × 18:2LPC%+7.43 × 18:1LPC%+0.024 × saturated LPC total amount=14.58].
The structure of the kit of two, examination colorectal cancer
1) set up comparison card
Comparison card A is made up of formula (straight line 1), horizontal ordinate 1 and the ordinate 1 of model 1;
Horizontal ordinate 1 is 0.023 × saturated LPC total amount+5.96 × 18:2LPC%;
Ordinate 1 is SPC total amount;
Saturated LPC total amount is the summation of 14:0LPC, 16:0LPC, 18:0LPC, 20:0LPC and 22:0LPC content;
Comparison card B is made up of formula (straight line 2), horizontal ordinate 2 and the ordinate 2 of model 2;
Horizontal ordinate 2 is 11.406 × 18:2LPC%+7.43 × 18:1LPC%;
Ordinate 2 is saturated LPC total amount;
Unsaturated LPC total amount is the summation of 18:2LPC, 18:1LPC, 20:4LPC and 22:6LPC content;
Saturated LPC total amount is the summation of 14:0LPC, 16:0LPC, 18:0LPC, 20:0LPC and 22:0LPC content;
18:1LPC% is the number percent that 18:1LPC content accounts for unsaturated LPC total amount.
18:2LPC% is the number percent that 18:2LPC content accounts for unsaturated LPC total amount;
2) structure of kit
Kit 1 comprises comparison card 1 or comparison card 2 and LC-MS instrument Applied Biosystems Sciex 3200QtrapTM massspectrometer, Applied Biosystems Sciex, Ontario, Canada.
Kit 2 comprises comparison card A, comparison card B and LC-MS instrument Applied Biosystems Sciex 3200QtrapTM massspectrometer, Applied Biosystems Sciex, Ontario, Canada.
The application of the kit of embodiment 2, examination colorectal cancer
Sensitivity and the specific detection of the kit (model 2) of 1, examination colorectal cancer
Get the blood of 120 Healthy Peoples contrast (all clinical identification) and 120 routine colorectal cancer patients (all clinical identification is made a definite diagnosis); According to embodiment 1 one 1 2) method collect that in vitro whole blood, lipid extraction method 1 extract, LC-MS instrument detects 18:1LPC, 18:2LPC, SPC total amount, 14:0LPC, 16:0LPC, 18:0LPC, 20:0LPC and 22:0LPC content;
Taking 11.406 × 18:2LPC%+7.43 × 18:1LPC% as horizontal ordinate, taking saturated LPC total amount as ordinate, make straight line with the formula [11.41 × 18:2LPC%+7.43 × 18:1LPC%+0.024 × saturated LPC total amount=14.58] of model 2, map.
Result is as shown in the right figure of Fig. 2, for (11.406 × 18:2LPC%+7.43 × 18:1LPC%) in the blood plasma of each colorectal cancer patients (C) and normal control (H) is to the mapping of saturated LPC amount, figure cathetus is: [11.406 × 18:2LPC%+7.43 × 18:1LPC%+0.024 × saturated LPC total amount=14.58], it is colorectal cancer patients that straight line downside is determined, and upside is judged to be normal healthy controls.18:2LPC% in formula and 18:1LPC% scope are 0-100%, represent that 18:2LPC or 18:1LPC account for respectively the number percent of all unsaturated LPC.
By analysis, have 116 samples at straight line downside, wherein, 100 samples derive from colorectal cancer patients (120), and 16 samples derive from normal healthy controls (120); In addition, 100 patients in 120 colorectal cancer patients by correct diagnosis (all at the downside of straight line, and survey and make a definite diagnosis through intestines microscopy), comprising 95 patients (55%) that T2 is interim, 58 50 patients (86%) and 35 29 patients (83%) that T4 is interim that T3 is interim, in 20 cancer patients that are missed, there is 1 to be the T0-1 phase, 4 T2 phases, 8 T3 phases and 6 T4 phases.
Therefore, sensitivity and specificity (the number 100/ tumor patient sum 120=83% being diagnosed in tumor patient) and 86% (number 100/ that is diagnosed as actual tumor patient in tumor patient is diagnosed as the number 116=86% of tumor patient) that be respectively 83%.
The serve as reasons ROC curve of the normal and colorectal cancer patients of differentiation that right figure sets up of the left figure of Fig. 2.
Sensitivity and the specific detection of the kit (model 1) of 2, examination colorectal cancer
Identical with above-mentioned 1 method, the sample detecting is also identical, taking 0.023 × saturated LPC total amount+5.96 × 18:2LPC% as horizontal ordinate, taking SPC total amount as horizontal ordinate, formula [0.023 × saturated LPC total amount+5.96 × 18:2LPC%+17.06 × SPC total amount=9.66] with model 1 is made straight line, maps.
Result is as shown in the right figure of Fig. 3, in blood plasma for each CRC patient (C) and normal control (H), (0.023 × saturated LPC total amount+5.96 × 18:2LPC%) maps to SPC total amount, figure cathetus is: [0.023 × saturated LPC total amount+5.96 × 18:2LPC%+17.06 × SPC total amount=9.66], it is colorectal cancer patients that straight line left side is determined, and right side is judged to be normal healthy controls.18:2LPC% scope in formula is 0-100%, represents that 18:2LPC accounts for the number percent of all unsaturated LPC.
By analysis, have 132 samples in straight line left side, wherein, 106 samples come from colorectal cancer patients (120), and 26 samples derive from normal healthy controls (120); In addition, have 106 patients by correct diagnosis (all in the left side of straight line, and through clinical detection, being really diagnosed as colorectal cancer) in 120 colorectal cancer patients, undetected patient is 14.
Therefore, sensitivity and specificity (the number 106/ tumor patient sum 120=88.3% being diagnosed in tumor patient) and 80% (number 106/ that is diagnosed as actual tumor patient in tumor patient is diagnosed as the number 132=80% of tumor patient) that be respectively 88.3%.
The serve as reasons ROC curve of the normal and colorectal cancer patients of differentiation that right figure sets up of the left figure of Fig. 3.
As can be seen from the above, LPC can be used as only one China crowd and ties the molecular labeling of rectum early diagnosis, also utilizes SPC to coordinate LPC to set up the model of distinguishing colorectal cancer and normal control, and model all has higher sensitivity and specificity.
To sum up, find that a series of phospholipid molecules that contain choline group can be used as the molecular labeling of diagnosis of colorectal carcinoma, utilize different phospholipid molecules and hypotype thereof can distinguish normal control/colorectal cancer,
The above-mentioned blood plasma of having collected 120 routine normal controls and 120 routine colorectal cancer patients in Chinese population, collection process is strictly regulated unification, and all experimenters accepted Sigmoidoscope and detected, and result and method of the present invention, without significant difference, illustrate that method of the present invention is correct.
Set up the molecular model of distinguishing Chinese population normal control and colorectal cancer patients, sensitivity and specificity can reach respectively 88% and 80% (or 83% and 86%).Utilize these lipid molecules the differentiation effect of colorectal cancer and normal control to be better than much studying in the last few years the mark of colorectal cancer early diagnosis.
According to above-mentioned two model testing results, can more accurately determine whether sample is colorectal cancer.
Claims (9)
1. a kit for examination or auxiliary examination colorectal cancer, comprises that the comparison of setting up for detection of the device of content of phospholipid with by described content of phospholipid blocks;
Described phosphatide is SPC, 14:0LPC, 16:0LPC, 18:2LPC, 18:1LPC, 18:0LPC, 20:4LPC, 20:0LPC, 22:6LPC and 22:0LPC;
The described comparison card of being set up by content of phospholipid is following 1) or 2);
1) be that contrast card A and comparison block any one in B;
2) contrast card A and comparison card B;
Described comparison card A is made up of straight line 1, horizontal ordinate 1 and ordinate 1;
Described straight line 1 is 0.023 × saturated LPC total amount+5.96 × 18:2LPC%+17.06 × SPC total amount=9.66;
Described horizontal ordinate 1 is 0.023 × saturated LPC total amount+5.96 × 18:2LPC%;
Described ordinate 1 is SPC total amount;
Described comparison card B is made up of straight line 2, horizontal ordinate 2 and ordinate 2;
Described straight line 2 is 11.41 × 18:2LPC%+7.43 × 18:1LPC%+0.024 × saturated LPC total amount=14.58;
Described horizontal ordinate 2 is 11.41 × 18:2LPC%+7.43 × 18:1LPC%;
Described ordinate 2 is saturated LPC total amount;
Described 18:2LPC% is the number percent that 18:2LPC content accounts for unsaturated LPC total amount;
Described 18:1LPC% is the number percent that 18:1LPC content accounts for unsaturated LPC total amount.
2. kit according to claim 1, is characterized in that:
Described saturated LPC total amount is the summation of 14:0LPC, 16:0LPC, 18:0LPC, 20:0LPC and 22:0LPC content;
Described unsaturated LPC total amount is the summation of 18:2LPC, 18:1LPC, 20:4LPC and 22:6LPC content.
3. kit according to claim 1 and 2, is characterized in that: the described device for detection of content of phospholipid is LC-MS instrument.
4. kit according to claim 3, is characterized in that:
The mobile phase that described detection adopts is that volume ratio is methyl alcohol, water, the ammonia water mixture of 90:10:0.1;
Described content of phospholipid is content of phospholipid in vitro blood.
5. kit according to claim 1 and 2, is characterized in that:
Described kit is made up of the described device for detection of content of phospholipid and the described comparison card of being set up by described content of phospholipid.
6. the application of arbitrary described kit in the product of preparation detection sample to be tested colorectal cancer in claim 1-5.
7. application according to claim 6, is characterized in that: described sample to be tested derives from people.
8. application according to claim 7, is characterized in that: described people is Asian.
9. application according to claim 8, is characterized in that: described people from Asia is Chinese.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210003024.3A CN102565267B (en) | 2012-01-06 | 2012-01-06 | Kit for screening colorectal cancer |
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| CN106893777B (en) * | 2017-03-02 | 2021-04-30 | 武汉艾米森生命科技有限公司 | Multi-site methylation kit for detecting colorectal cancer related genes and application thereof |
| CN114544826B (en) * | 2020-11-24 | 2023-12-08 | 重庆医科大学 | Application of reagent for detecting histidine in blood plasma in preparation of depression detection kit |
| CN114544822B (en) * | 2020-11-24 | 2023-10-24 | 重庆医科大学 | Application of reagent for detecting lysophosphatidylcholine (22:0) in blood plasma in preparation of depression detection kit |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101498700A (en) * | 2009-02-16 | 2009-08-05 | 桑建利 | Production and use of reagent kit for measurement of lipide molecule content in human blood sample and colorectal carcinoma diagnosis |
| CN101688239A (en) * | 2007-02-12 | 2010-03-31 | 约翰·霍普金斯大学 | The early detection of colorectal carcinoma and prognosis |
| CN101802616A (en) * | 2007-07-19 | 2010-08-11 | 生物梅里埃公司 | The liver fatty acid-binding protein of in-vitro diagnosis colorectal cancer, CEA and CA19-9 assay method |
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| FR2944019B1 (en) * | 2009-04-03 | 2011-04-22 | Biomerieux Sa | METHOD FOR DETERMINING PRODEFENSIN-A6 FOR IN VITRO DIAGNOSIS OF COLORECTAL CANCER |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101688239A (en) * | 2007-02-12 | 2010-03-31 | 约翰·霍普金斯大学 | The early detection of colorectal carcinoma and prognosis |
| CN101802616A (en) * | 2007-07-19 | 2010-08-11 | 生物梅里埃公司 | The liver fatty acid-binding protein of in-vitro diagnosis colorectal cancer, CEA and CA19-9 assay method |
| CN101498700A (en) * | 2009-02-16 | 2009-08-05 | 桑建利 | Production and use of reagent kit for measurement of lipide molecule content in human blood sample and colorectal carcinoma diagnosis |
Non-Patent Citations (8)
| Title |
|---|
| Electrospray Ionization Mass Spectrometry Analysis of Lysophospholipids in Human Ascitic Fluids:Comparison of the Lysophospholipid Contents in Malignant vs Nonmalignant Ascitic Fluids;Yi-jin Xiao et al;《Analytical Biochemistry》;20010315;第290卷(第2期);第302-313页 * |
| Evaluation of Plasma Lysophospholipids for Diagnostic Significance Using Electrospray Ionization Mass Spectrometry (ESI-MS) Analyses;YIJIN XIAO et al;《LYSOPHOSPHOLIPIDS AND EICOSANOIDS IN BIOLOGY AND PATHOPHYSIOLOGY》;20001231;第905卷;第242-259页 * |
| Yi-jin Xiao et al.Electrospray Ionization Mass Spectrometry Analysis of Lysophospholipids in Human Ascitic Fluids:Comparison of the Lysophospholipid Contents in Malignant vs Nonmalignant Ascitic Fluids.《Analytical Biochemistry》.2001,第290卷(第2期), * |
| YIJIN XIAO et al.Evaluation of Plasma Lysophospholipids for Diagnostic Significance Using Electrospray Ionization Mass Spectrometry (ESI-MS) Analyses.《LYSOPHOSPHOLIPIDS AND EICOSANOIDS IN BIOLOGY AND PATHOPHYSIOLOGY》.2000,第905卷 * |
| 乔小放.血浆溶血磷脂的检测在胃癌早期诊断中的应用.《医药卫生科技辑》.2008,第E072-96页. * |
| 血浆溶血磷脂的检测在胃癌早期诊断中的应用;乔小放;《医药卫生科技辑》;20081115;第E072-96页 * |
| 赵素敏等.赵素敏等.《色谱》.2011,第29卷(第9期), * |
| 赵素敏等;赵素敏等;《色谱》;20110930;第29卷(第9期);第843-850页 * |
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