CN102565267A

CN102565267A - Kit for screening colorectal cancer

Info

Publication number: CN102565267A
Application number: CN2012100030243A
Authority: CN
Inventors: 张俊杰; 李颂; 宋建稳; 徐燕; 桑建利; 丛羽生
Original assignee: Beijing Normal University
Current assignee: Beijing Normal University
Priority date: 2012-01-06
Filing date: 2012-01-06
Publication date: 2012-07-11
Anticipated expiration: 2032-01-06
Also published as: CN102565267B

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

The kit of examination colorectal cancer

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, surpasses the million people in the last few years every year and dies from colorectal cancer.Because the 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 that 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 present colorectal cancer early detection rate less than 40%, in case shift colorectal cancer patients survival rate less than 10%." goldstandard " of current diagnosis colorectal cancer is Sigmoidoscope, generally acknowledge at present, the age greater than 50 years old should accept the examination of intestines mirror per capita.Authoritative department such as the U.S. and Europe points out, for have familial medical history occurred frequently or since 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 in health check-up, extensively carrying out the survey of intestines microscopy at present, and experimenter's misery is also self-evident.In China's health check-up, extensively carrying out Sigmoidoscope at present detects unrealistic.Therefore, colorectal cancer early diagnosis molecular labeling becomes and studies one of focus now in the screening blood plasma.

People have confirmed to be present in a large number the biomarker in colon, ight soil or the serum; Like carcinomebryonic antigen (CEA) etc.; Be used for the early diagnosis and the prediction of colon cancer, but the sensitivity of the screening method of great majority recommendation at present and specificity are all below 50%.Recently, also reported in some tissues in succession, the colorectal cancer molecular labeling in the ight soil and in the serum, but its effect is also far below Sigmoidoscope.For example, if serve as a mark with the special site that methylates of faeces DNA, when specificity reached 90%, sensitivity had only 50%.And this detection easy to detect not as blood marker certainly, therefore at present more medical personnel hopes to develop the mark of diagnosis of colorectal carcinoma in the blood.The research report is arranged, and serum colon cancer specific antigen CCSA-3 and CCSA-4 possibly become the mark of diagnosis of colorectal carcinoma, and it detects effect and also need confirm.Annual approximately report has thousands of kinds of tumour molecular marked compounds, but being applied to clinical molecular marker does not at last almost have, and the defective that their exist concentrates on sample size less (＜30), not in not agnate or independent experiment by certain etc.More crucial is that the disunity of standard is difficult to promoted by these molecular markers in sample collection, pre-service and target molecule extractive process.

To sum up, higher owing to intestines mirror expense, painful big and operation requires high reason to reduce the degree that patient would like to do this inspection, causes a lot of patients to be dragged to and has become incurable disease late period.Therefore need develop a kind of simple and reliable, big flux inspection method of not having wound or Wicresoft that can be applicable to the colorectal cancer early screening in a hurry.

The lysophosphatide molecule of biologically active causes scientist's concern day by day 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 take place and be adsorbed in multiple activity.Therefore these lysophosphatide molecules can be participated in angiogenesis, wound healing, immunity, atherosclerotic generation and the adjusting that becomes 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 through ATX catalysis, and S1P and LPA are very remarkable to the regulatory function of tumour as the 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 maybe be as the biomarker of early diagnosis of tumor.Based on lipid molecule, the particularly important physiological of biologically active lipid molecule and pathology sense have proposed the notion of lipid group and function lipid group in the world recently.Except that existing genome and proteomics, the research and development of lipid group will play an important role in the early diagnosis and therapy of tumour.In addition, can from people's body fluid (serum), directly detecting unusually of early stage biologically active lipid molecule metabolism takes place in tumour.By comparison, traditional tissue samples detects that not only wound is big, and tumour is difficult to accomplish early diagnosis.Compare with protein marker with gene, as the metabolic marker thing, the detection of biologically active lipid molecule is more convenient, quick, is suitable for clinical expansion.Therefore, set up sensitive, biologically active lipid analysis method accurately and efficiently, screening has great importance to early diagnosis of tumor to different TS biologically active lipid molecule labels.

Be different from protein and polypeptide, exploitation is directed to simple lysophosphatide molecule, and is very difficult like 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 are once by the analyzing and testing that is applied to lysophosphatide of success, like thin-layer chromatography (TLC), and phosphorous determination and gas chromatography (GC) etc.But the analytic process that has in these methods is complicated, and workload is big, the then poor sensitivity that has, and the low lysophosphatide of content in the analysing body fluid efficiently, and have no a kind of method can simply be used to separate and analyze the hypotype of lysophosphatide.

Summary of the invention

The kit that the purpose of this invention is to provide a kind of examination or auxiliary examination colorectal cancer.

The kit of examination provided by the invention or auxiliary examination colorectal cancer comprises being used to detect the device of content of phospholipid and the comparison card of being set up by said content of phospholipid;

Said phosphatide is SPC, 14:0LPC, 16:0LPC, 18:2LPC, 18:1LPC, 18:0LPC, 20:4LPC, 20:0LPC, 22:6LPC and 22:0LPC.

The said 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 among the B;

2) comparison card A and comparison card B;

Said comparison card A is made up of straight line 1, horizontal ordinate 1 and ordinate 1;

Said straight line 1 is 0.023 * saturated LPC total amount+5.96 * 18:2LPC%+17.06 * SPC total amount=9.66;

Said horizontal ordinate 1 is 0.023 * saturated LPC total amount+5.96 * 18:2LPC%;

Said ordinate 1 is SPC total amount (content);

Above-mentioned straight line 1 is the linear function of ordinate 1 about horizontal ordinate 1;

Said comparison card B is made up of straight line 2, horizontal ordinate 2 and ordinate 2;

Said straight line 2 is 11.41 * 18:2LPC%+7.43 * 18:1LPC%+0.024 * saturated LPC total amount=14.58;

Said horizontal ordinate 2 is 11.406 * 18:2LPC%+7.43 * 18:1LPC%;

Said ordinate 2 is saturated LPC total amount;

Above-mentioned straight line 2 is the linear function of ordinate 2 about horizontal ordinate 2;

Said 18:2LPC% accounts for the number percent of unsaturated LPC total amount for 18:2LPC content;

Said 18:1LPC% accounts for the number percent of unsaturated LPC total amount for 18:1LPC content.

Said saturated LPC total amount is the summation of 14:0LPC, 16:0LPC, 18:0LPC, 20:0LPC and 22:0LPC content;

Said 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 said device that is used to detect content of phospholipid is the LC-MS appearance.

Said LC-MS appearance is Applied Bio systems S ciex 3200Qtrap ^TMMass spectrometer, Applied Biosystems Sciex, Ontario, Canada, it detects used moving phase is that volume ratio is 90: 10: 0.1 methyl alcohol, water, an ammonia water mixture;

The flow velocity of the moving phase that said detection is adopted is 0.2ml/min,

Do not need pillar when detecting LPC, SPC, L-PAF and each subclass of SM; 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).

Said content of phospholipid is content of phospholipid in the blood that exsomatizes.

Said kit is made up of said device and the said comparison card of being set up by said content of phospholipid that is used to detect content of phospholipid.

The application of described kit in the product of preparation detection sample to be tested colorectal cancer also is the scope that the present invention protects.

Said sample to be tested derives from the people, specifically derives from the Asian, especially is preferably Chinese.

Experiment of the present invention proves; The present invention's in Chinese population (each 120 example of normal control and tumor patient) has furtherd investigate content (LPC, LPA, the SPC of a series of phospholipid molecules; S1P; SM and haemolysis platelet factor L-PAF), particularly contain the content of lipid molecule (LPC, SM and SPC) various subclass in blood plasma of choline group, deeply probe into the variation of these lipids in colorectal cancer; And then develop effective colorectal cancer early diagnosis molecular labeling; Set up the Molecular Detection model, sensitivity that detects according to this model and specificity can reach 88% and 80% (perhaps 83% and 86%), utilize these lipid molecules the differentiation effect of colorectal cancer and normal control to be superior to much studying in the last few years the mark of colorectal cancer early diagnosis.

In the screening of the present invention, be the basis, developed the technology of lysophosphatide molecule in the detection by quantitative human plasma with Electro-spray ionization mass spectrometry (ESI-MS, triple level Four tandem mass spectrometers).The advantage of Mass Spectrometer Method is: (1) highly sensitive, good reproducibility; (2) it not only can detect a large amount of different types of lipid molecules simultaneously, and can differentiate the different subtype with a kind of lipid molecule; (3) detect plasma sample and have advantages such as sampling is convenient, wound is little, and required sample size has only 10-50 μ l; (4) utilize a self-actuated sampler, the very robotization that just can become of this detection technique.Therefore, this detection method has very big potentiality in clinical practice.Use correlation technique, 2007, the Xu Yan of indiana ,US university professor reported that at first LPC can be used as the colorectal cancer early diagnosis marker of American population in the laboratory.But for the early diagnosis of cancer molecular labeling, the independent detection of striding race and crowd is most important, and 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 the 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 every year and surpass 100; 000 piece of article about " cancer marker " is delivered; But these molecular markers are applied to the few of clinical examination at last; There is many difference in the different experiments chamber to the testing result of same mark, and the detection method of the difference of sample collection mode difference, extracting mode and complicacy etc. all is the reason that these marks are difficult to be employed.And of the present invention these are contained the lipid molecule testing process of choline group; Take a sample, extract and detect highly standardized; And utilize tens microlitre blood plasma and the simple organic reagent just can the accurately quantitative content of these lipid molecules in blood plasma; Can carry out in the routine physical examination, and, refer to that promptly the blood chemical examination just might provide preliminary foundation for diagnosis of colorectal carcinoma its screening indexes as the colorectal cancer early diagnosis.

Description of drawings

Fig. 1 is saturated LPC, unsaturated LPC, total LPC and (16:0+18:0) the changes of contents trend of SM content in normal control and colorectal carcinoma patient body

Fig. 2 distinguishes normal control and colorectal cancer patients model in the Chinese population for utilizing LPC

Fig. 3 distinguishes normal control and colorectal cancer patients model in the Chinese population for utilizing LPC and SPC

Embodiment

Employed experimental technique is conventional method like no specified otherwise among the following embodiment.

Used material, reagent etc. like no specified otherwise, all can obtain from commercial sources among the following embodiment.

The structure of the kit of embodiment 1, examination colorectal cancer

One, the foundation of molecular model

1, blood collecting, lipid molecule extract and detect

1) research object

All sample collections are in May, 2007-2010 year May, and sample is from the Second Artillery Force of entire PLA hospital general, Beijing Hospital, second affiliated hospital of Peking University.Getting morning on an empty stomach, patient blood carries out pre-service.This project is examined all subjects signed Informed Consent Forms through Institutional Review Board.

Table 1 has been summarized the basic condition of research object: tried that masculinity proportion is respectively 68% and 59% in healthy sample and the tumor sample; 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 rectal neoplasm mostly.Tumour above 75% is T3/T4 phase (93/120), and only about half of tumour is N0 (58/120,48%).See that 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 collecting, lipid molecule extract and detect

In containing the pipe of EDTA, collect above-mentioned 1) the stripped whole blood of sample, room temperature (25 ℃) 1, centrifugal 15 minutes of 750g, with pack into the pipe of silicidation of the blood plasma branch that exsomatizes ,-80 degree refrigerators are preserved, and avoid multigelation.

The extraction of LPC, SPC, SM and L-PAF (lipid extraction method 1): in the glass centrifuge tube, add 0.5ml PBS (Hyclone company; SH30256.01B); (avanti polar lipids 855475P 100mM), adds 3ml methyl alcohol/chloroform (volume ratio is 2: 1) to add 100 μ l 12:0LPC.In above-mentioned solution, add the above-mentioned stripped blood plasma that obtains of 10 μ l, placed on ice behind the whirlpool mixing 10 minutes.Add the 1ml chloroform, 1.3ml distilled water, mixing, (25 ℃) 1 are mixed in the chamber, and centrifugal 10 minutes of 750g takes off a layer organic phase, and nitrogen dries up, and with 1ml methanol (9: 1) dissolving, gets machine testing LPC, SPC, SM and L-PAF and each subclass content thereof on the 200 μ l.

The extraction of LPA and S1P (lipid extraction method 2): in the glass centrifuge tube, add 0.5ml PBS, 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, placed on ice behind the whirlpool mixing 10 minutes.Add the 1ml chloroform, 1.3ml distilled water, mixing, room temperature 1, centrifugal 10 minutes of 750g takes off a layer organic phase, and nitrogen dries up, and with the dissolving of 150 μ l methanol (9: 1), gets machine testing LPA and S1P and each subclass content thereof on the 150 μ l.

The above-mentioned machine testing of going up all adopts the LC-MS system: LC-MS appearance Applied Biosystems Sciex 3200QtrapTM massspectrometer; Applied Biosystems Sciex; Ontario, Cahada, moving phase is: methanol/ammoniacal liquor (90: 10: 0.1); The flow velocity of said detection content of phospholipid moving phase is 0.2ml/min; Measure LPC, SPC, L-PAF and each subclass of SM and do not need pillar; The pillar model of measuring LPA and 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 available from Avanti Polar Lipids company (seeing 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	Synthetic by the said firm
		22:0LPC	855779C
22:6LPC	Synthetic 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 influence of age to content of phospholipid in the blood plasma that detects

Generally acknowledge that at present the morbidity of colorectal cancer is directly related with the age, the age is big more, and incidence probability is high more, if but with the age as colorectal cancer early diagnosis label, sensitivity has 79.8%, but specificity has only 23.9%.And all lipid variation tendencies and age that the present invention detects do not have direct relation, so these lipid molecules that contain the choline group are diagnostic markers separate with the age.

2) content and the relation between sex of phospholipid molecule in blood plasma

Testing result is as described in Table 4, can find out, if carry out the t check; With p=0.01 as standard; Male sex 18:2LPC, 18:1LPC and the total amount of unsaturated LPC in blood plasma be a little more than the women, and difference is also not obvious, so the content of these phospholipid molecules and sex do not have obvious relation yet.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 the blood plasma.

Detected 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.

The normal healthy controls group is compared with the cancer group, and 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 all significantly is lower than normal control group (table 5).Saturated LPC, unsaturated LPC and total LPC content all significantly descend 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 might become molecular labeling shared in not agnate or the 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).Content and the stage of tumor and the location independent (data now shown) of all phospholipid molecules that detect.Detect the difference in the 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) sets up the 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 the 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;

18:1LPC% accounts for the number percent of unsaturated LPC total amount for 18:1LPC content.

18:2LPC% accounts for the number percent of unsaturated LPC total amount for 18:2LPC content;

2) structure of kit

Kit 1 comprises comparison card 1 or comparison card 2 and LC-MS appearance Applied Biosystems Sciex 3200QtrapTM massspectrometer, Applied Biosystems Sciex, Ontario, Canada.

Kit 2 comprises comparison card A, comparison card B and LC-MS appearance Applied Biosystems Sciex 3200QtrapTM massspectrometer, Applied Biosystems Sciex, Ontario, Canada.

The application of the kit of embodiment 2, examination colorectal cancer

The sensitivity and the specific detection of the kit of 1, examination colorectal cancer (model 2)

Get the blood of 120 healthy subjects 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 stripped whole blood, lipid extraction method 1 extract, the LC-MS appearance detects 18:1LPC, 18:2LPC, SPC total amount, 14:0LPC, 16:0LPC, 18:0LPC, 20:0LPC and 22:0LPC content;

With 11.406 * 18:2LPC%+7.43 * 18:1LPC% is horizontal ordinate, is ordinate with saturated LPC total amount, makes straight line with the formula [11.41 * 18:2LPC%+7.43 * 18:1LPC%+0.024 * saturated LPC total amount=14.58] of model 2, maps.

The result is shown in the right figure of Fig. 2; In the blood plasma for each colorectal cancer patients (C) and normal control (H) (11.406 * 18:2LPC%+7.43 * 18:1LPC%) to the mapping of saturated LPC amount; The figure cathetus is: [11.406 * 18:2LPC%+7.43 * 18:1LPC%+0.024 * saturated LPC total amount=14.58]; The straight line downside is a colorectal cancer patients by judgement, and upside is judged to be normal healthy controls.18:2LPC% in the formula and 18:1LPC% scope are 0-100%, represent 18:2LPC or 18:1LPC to account for the number percent of all unsaturated LPC respectively.

Through analyzing, have 116 samples at the 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 are by correct diagnosis (survey is made a definite diagnosis all at the downside of straight line, and through the 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; There is 1 to be the T0-1 phase by among 20 cancer patients that omit, 4 T2 phases, 8 T3 phases and 6 T4 phases.

Therefore, sensitivity and specificity (the number 100/ tumor patient sum 120=83% that is diagnosed in the tumor patient) and 86% (number 100/ that is diagnosed as actual tumor patient in the 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.

The sensitivity and the specific detection of the kit of 2, examination colorectal cancer (model 1)

Identical with above-mentioned 1 method; The sample that detects is also identical; With 0.023 * saturated LPC total amount+5.96 * 18:2LPC% is horizontal ordinate; With the SPC total amount is horizontal ordinate, makes straight line with the formula [0.023 * saturated LPC total amount+5.96 * 18:2LPC%+17.06 * SPC total amount=9.66] of model 1, maps.

The result is shown in the right figure of Fig. 3; (map to the SPC total amount in 0.023 * saturated LPC total amount+5.96 * 18:2LPC%) in the blood plasma for each CRC patient (C) and normal control (H); The figure cathetus is: [0.023 * saturated LPC total amount+5.96 * 18:2LPC%+17.06 * SPC total amount=9.66]; The straight line left side is a colorectal cancer patients by judging, the right side is judged to be normal healthy controls.18:2LPC% scope in the formula is 0-100%, represents 18:2LPC to account for the number percent of all unsaturated LPC.

Through analyzing, have 132 samples in the straight line left side, wherein, 106 samples come from colorectal cancer patients (120), and 26 samples derive from normal healthy controls (120); In addition, 106 patients are arranged by correct diagnosis (all in the left side of straight line, and through Clinical detection, being diagnosed as colorectal cancer really) in 120 colorectal cancer patients, the patient of omission is 14.

Therefore, sensitivity and specificity (the number 106/ tumor patient sum 120=88.3% that is diagnosed in the tumor patient) and 80% (number 106/ that is diagnosed as actual tumor patient in the 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.

Can find out that from above-mentioned LPC can be used as the molecular labeling that the crowd of a Chinese ties the rectum early diagnosis, also utilize SPC to cooperate LPC to set up the model of distinguishing colorectal cancer and normal control, model all has higher sensitivity and specificity.

To sum up, a series of phospholipid molecules of finding to contain the 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 the Chinese population; Collection process is strictly regulated unification; And all experimenters accepted Sigmoidoscope and detected, and result and method of the present invention do not have significant difference, explain 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 88% and 80% respectively (perhaps 83% and 86%).Utilize these lipid molecules the differentiation effect of colorectal cancer and normal control to be superior to much studying in the last few years the mark of colorectal cancer early diagnosis.

According to above-mentioned two model testing results, can confirm more accurately whether sample is colorectal cancer.

Claims

1. the kit of an examination or auxiliary examination colorectal cancer comprises being used to detect the device of content of phospholipid and the comparison card of being set up by said content of phospholipid;

2. kit according to claim 1 and 2 is characterized in that:

The said 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 among the B;

2) contrast card A and comparison card B;

Said ordinate 1 is the SPC total amount;

Said horizontal ordinate 2 is 11.406 * 18:2LPC%+7.43 * 18:1LPC%;

Said ordinate 2 is saturated LPC total amount;

3. kit according to claim 1 and 2 is characterized in that:

Said unsaturated LPC total amount is the summation of 18:2LPC, 18:1LPC, 20:4LPC and 22:6LPC content.

4. according to arbitrary described kit among the claim 1-3, it is characterized in that:

5. according to arbitrary described kit among the claim 1-4, it is characterized in that:

The moving phase that said detection is adopted is that volume ratio is 90: 10: 0.1 methyl alcohol, water, an ammonia water mixture;

6. according to arbitrary described kit among the claim 1-5, it is characterized in that:

7. the application of the kit described in the claim 1-6 in the product of preparation detection sample to be tested colorectal cancer.

8. application according to claim 7 is characterized in that: said sample to be tested derives from the people, specifically derives from the Asian, especially is preferably Chinese.