CN1553190A - Four mucin enzyme linked immunosorbent assay reagent box for early diagnosing oophoroma - Google Patents
Four mucin enzyme linked immunosorbent assay reagent box for early diagnosing oophoroma Download PDFInfo
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Abstract
A reagent kit is composed of standard products, multiple clone antibody encapsulation plate, enzyme labelling single clone antibody, quality control product and auxiliary reagent. The preparing method of the kit and inspection method are provided in the present invention.
Description
This present invention belongs to medical science and Biological Detection field, particularly, the present invention relates to a kind of enzyme-linked immunologic detecting kit that is used to diagnose early ovarian cancer, promptly the invention provides a kind of ovarian cancer protein mark TN (is Tetranectin, be called for short TN) detection kit, can be used for the detection of good, the pernicious diagnosis, particularly early ovarian cancer of ovarian neoplasm, and the detection of mucus epithelial ovarian cancer; The monitoring and the prognosis evaluation that also can be used for the ovarian cancer post operation chemotherapy effect.
Oophoroma is the malignant tumour that cure rate is minimum in the gynecological tumor, mortality ratio is the highest in worldwide always.At present the incidence of disease of oophoroma in China's gynecological cancer accounts for the 3rd and rise to first by the eighties.Oophoroma first visit patient more than 80% is arranged approximately to late period (III phase or IV phase), even operation at once, the postoperative five year survival rate is also only less than 15%, and the badness come-off of oophoroma operation prognosis has not almost improved since nearly 50 years.Because the pathogenesis of oophoroma is not still asked Chu so far, therefore be difficult to take proper prophylactic methods.In order to reduce the harm of oophoroma, key is still found in early days, early treatment.Though basis and clinical research have had been found that the tumor markers of some oophoromas, as carbohydrate antigen 125 (CA125), carcinomebryonic antigen (CEA), tissue polypeptide antigen (TPA), macrophage colony stimulatory factor (M-CSA) and inhibin etc., but really offer the also few of clinical use.Even it is still many with of the reference frame of CA125 single index in hospital with good conditionsi in recent years as ovarian cancer diagnosis, postoperative chemotherapy monitoring and prognosis evaluation.Because CA125 also is used for the detection of other malignant tumours, be not specific index therefore to oophoroma.And CA125 only has rising in the serous ovarian cancer patients serum, to the then not reaction of non-serous ovarian cancer, especially mucous ovarian cancer.CA125 is also insensitive to early ovarian cancer.CA125 obviously can not meet clinical needs as the unique index that detects oophoroma.Purpose of the present invention is used the deficiency of individual event CA125 as the ovarian cancer diagnosis index at present clinically in order to solve just, TN is used for the early diagnosis of oophoroma and the diagnosis of mucus epithelial ovarian cancer as a kind of new protein marker, and detection kit and assay method easy and simple to handle, accurate sensitivity are provided, thereby can improve the rate that picks of oophoroma, so that can take effective comprehensive therapeutic plan as early as possible clinically, reach the purpose that improves survival rate, reduces mortality ratio.
Therefore, the purpose of this invention is to provide a kind of enzyme-linked immunologic detecting kit that is used to diagnose early ovarian cancer.
Above-mentioned purpose of the present invention is implemented by following technical proposal:
A kind of ovarian cancer protein mark TN (is Tetranectin, be called for short TN) enzyme-linked immunologic detecting kit, it is characterized in that it is made up of by plate, horseradish peroxidase (HRP) mark polyclonal antibody, quality-control product and auxiliary reagent TN standard items, polyclonal antibody bag.
The method for making of kit of the present invention comprises that the preparation, polyclonal antibody bag of standard items are by the making of plate, enzyme mark Polyclonal Antibody Preparation, the preparation of quality-control product and the preparation of auxiliary reagent.
TN standard items in the kit of the present invention extract preparation from human plasma.A kind of preparation process comprises: collect normal plasma (being negative patient through mensuration such as HIV, hepatitis virus, syphilis, AIDS), low tempertaure storage is standby; Above-mentioned blood plasma carries out the separation and purification of TN through saturated ammonium sulphate, ion-exchange chromatography and three steps of affinity chromatography, two step of the back order of chromatographies can conversion, and a kind of TN purification sequence that kit of the present invention uses is followed successively by saturated ammonium sulphate, ion-exchange chromatography and affinity chromatography; The pure product of TN that obtain are diluted by the typical curve peak concentration of kit, carry out packing, freeze drying then, promptly obtain the TN standard items.
The polyclonal antibody coated elisa plate that polyclonal antibody bag in the kit of the present invention can be obtained the immunity of rabbit or mouse with the pure product of TN by plate and making.Polyclonal antibody bag in the kit of the present invention is made of the mouse-anti people polyclonal antibody coated elisa plate that the pure product of TN obtain mouse immune by plate.ELISA Plate can be selected homemade plate or import plate for use; Specification can be dull and stereotyped or 12 * 8,6 * 8 removable battens in 96 holes.Kit of the present invention adopts a kind of import ELISA Plate (Costar company product).A kind of making step of polyclonal antibody coated elisa plate is as follows:
A) prepare polyclonal antibody: after routinely the Balb/c mouse being carried out immunity with the pure product of TN, again at intraperitoneal injection murine myeloma cell SP2/0; Collect ascites and behind reorganization Protein G prepackage chromatographic column purifying, promptly obtain the TN polyclonal antibody;
B) bag quilt: above-mentioned polyclonal antibody is added each hole of ELISA Plate with 0.05M carbonate buffer solution dilution back, every hole 100ul, absorption is spent the night, wash plate with the tween phosphate buffer, spend the night with the sealing of tween phosphate buffer again, dry after the drying, promptly obtain the polyclonal antibody coated elisa plate.
Enzyme mark polyclonal antibody in the kit of the present invention prepares with horseradish peroxidase (HRP) mark TN polyclonal antibody.A kind of enzyme mark Polyclonal Antibody Preparation step is as follows:
A) in the good TN antibody-solutions of purifying, add SATA solution, room temperature reaction 0.5 hour;
B) add deacetylation solution, incubated at room 2 hours;
C) separate acetylation SATA derivant (promptly many anti-) and secondary product (oxammonium hydrochloride) with desalting column;
D) with above-mentioned how anti-solution with the HRP of maleic amide (Maleimide) activation room temperature reaction 1 hour, promptly obtain HRP enzyme mark polyclonal antibody.
Quality-control product in the kit of the present invention is to use the PHS, requires to dilute and allocates preparation according to the standard curve range of this kit.A kind of quality-control product preparation method that kit of the present invention adopts comprises the steps:
A) collect normal human serum (being negative patient), in-20 ℃ of preservations through mensuration such as HIV, liver viroid, AIDS, syphilis;
Each serum that b) will accumulate mixes, and measures TN concentration with the ELISA method, and by the typical curve requirement, and the concentration of two parts of pooled serums is transferred to 10 ± 2ng/ml, 50 ± 10ng/ml scope respectively;
C) with the requirement packing by each kit of two parts of serum that step b) obtained, freeze drying promptly is prepared into low, high value quality-control product.
Auxiliary reagent in the kit of the present invention comprises the integrated enzyme reaction substrate solution, colour developing liquid, reaction terminating liquid and cleaning buffer solution.A kind of method of preparing auxiliary reagent is as follows:
A) substrate solution: 3% superoxol of phosphoric acid-citrate buffer solution (pH5.0) preparation;
B) colour developing liquid: tetramethyl benzidine (TMB) methanol solution, concentration is 0.1mg/ml;
C) reaction terminating liquid: 2M sulfuric acid
D) cleaning buffer solution (20 times of concentrates, 20X): 0.05% polysorbas20 solution of PBS (pH7.4) preparation.
The assay method of kit of the present invention is characterized in that it is undertaken by following step successively:
A) antigen-antibody reaction: in the micropore of the antibody sandwich plate that kit provides, add standard items, quality-control product that 100 μ l have diluted good variable concentrations respectively, or the test serum sample, 37 ℃ of water bath heat preservations 60 minutes; Use cleaning buffer solution (1X) to repeat to wash plate 4 times then.
B) integrated enzyme reaction: HRP-polyclonal antibody solution is added each hole, every hole 100 μ l, 37 ℃ of water bath heat preservations 60 minutes.Repeat to wash plate operation 4 times.
C) chromogenic reaction: every hole adds substrate solution, each 50 μ l of colour developing liquid successively, 37 ℃ of water bath heat preservations 10 minutes, and every hole adds 50 μ l reaction terminating liquids again and finishes reaction.
D) colorimetric:, measure OD value and record at 450nm with microplate reader with the light absorption value zeroing in blank hole.
E) result calculates:
A. production standard curve: with standard items concentration is horizontal ordinate, and the OD value that standard items are measured is an ordinate, makes typical curve; Basis of calculation curvilinear regression coefficients R
2, work as R
2This was measured effectively in＞0.98 o'clock;
B. pass judgment on quality-control product concentration: the OD value according to quality-control product, read corresponding concentration value from typical curve; When the concentration of low value quality-control product, high value quality-control product was all in given range, this mensuration was judged to effectively;
C. calculate the test serum sample concentration:, calculate the TN concentration of test serum sample from typical curve according to the OD value of sample to be tested when typical curve and quality-control product all are determined when effective.
Detect index CA125 with existing oophoroma and compare, kit of the present invention has the following advantages:
A) kit of the present invention has overcome the insensitivity of CA125 to early ovarian cancer with the diagnosis index of TN as early ovarian cancer, only has 50% patient's change of serum C A125 horizontal abnormality to raise in I phase ovarian cancer patients; And TN presents unusual decline in I phase～IV phase ovarian cancer patients.
B) CA125 can reach more than 80% the positive rate of picking of serosity epithelial ovarian cancer, yet to the not reaction of mucus epithelial ovarian cancer; Kit of the present invention has overcome individual event CA125 detection oophoroma and has had higher false-negative deficiency, and TN not only can pick the serosity epithelial ovarian cancer, and effective equally to the mucus epithelial ovarian cancer.TN uses separately or unites use with CA125, can improve the rate that picks of early ovarian cancer effectively, especially can improve the rate that picks of mucus epithelial ovarian cancer.
Further specify the present invention with embodiment below.It should be understood that embodiments of the invention are to be used to illustrate the present invention rather than limitation of the present invention.Essence according to the present invention all belongs to the scope of protection of present invention to the simple modifications that the present invention carries out.Except as otherwise noted, the percentage among the present invention is percent by weight.
Embodiment one: the application example of the inspection-free test agent box of the quantitative enzyme of TN method for making
The inspection-free test agent box of the quantitative enzyme of a kind of ovarian cancer protein mark TN (48 person-portion), its composition comprises:
1 bottle of TN freeze-drying standard items;
TN polyclonal antibody bag is by 1 of plate (48 hole);
1 bottle of the TN polyclonal antibody of horseradish peroxidase (HRP) mark, the 6ml/ bottle;
1 bottle of freeze-drying low value quality-control product;
1 bottle of the high value of freeze-drying quality-control product;
1 bottle of substrate solution, the 3ml/ bottle;
Colour developing liquid (TMB) 1 bottle, the 3ml/ bottle;
1 bottle of reaction terminating liquid, the 3ml/ bottle;
1 bottle of cleaning buffer solution (20X concentrates), the 15ml/ bottle.
Concrete operations are as follows:
1. prepare the TN standard items:
1) collect blood plasma: the human normal plasma that secures good health from hospital or blood station, standby in-70 ℃ of preservations;
2) separation and purification: use AKTA protein purification instrument (Pharmacia company product, model Purifier 100) separation and purification TN from above-mentioned blood plasma.The operation steps of selecting for use is followed successively by: a) saturated ammonium sulphate: add solid ammonium sulfate to 50% saturation degree in proportion earlier add equivalent PBS solution in blood plasma after, again pH is transferred to 5.8, the centrifugal albumen precipitation of collecting this moment; B) DEAE ion-exchange chromatography, damping fluid are 0.02M Tris-HCl, pH7.2,1M NaCl, flow velocity 3ml/min; C) Lysine affinity chromatography, damping fluid are 0.02M Tris-HCl, pH7.2,1M NaCl, flow velocity 1.0ml/min.Monitor with the concentration gradient of UNICRN V400 software control damping fluid and the uv absorption of protein peak in the chromatography process.The activity of each step protein peak that chromatography obtains detects the ELISA method that adopts.The activated protein eluting peak that with NaCl concentration is at last at 20% o'clock merges, and concentrates behind the dress bag filter dialysis desalination, be stored in after the packing-70 ℃ to be identified.
3) the pure product of TN by first required concentration packing of typical curve in the kit instructions (75ng/ml), freeze drying, be stored in 4 ℃.
2. make TN polyclonal antibody bag by plate:
1) TN polyclonal antibody preparation:
A) mouse immune: selecting body weight for use is 10 of the Balb/c male mices (available from Beijing Experimental Animal Center) of 20-22g, and every usefulness contains the physiological saline of the pure product of 50 μ g TN and the Freund's complete adjuvant of 2 times of volumes is made 0.5ml emulsion, is injected in the mouse peritoneal.The incomplete Freund that two Zhou Houyong contain the physiological saline of the pure product of 50 μ g TN and 2 times of volumes is made 0.5ml emulsion and is done booster shots; Repeat booster shots once after 2 weeks.Next day after the repetition booster shots, injection murine myeloma cell SP2/0 2 * 10 in every mouse peritoneal
6/ 0.5ml.One week back mouse generation ascites;
B) ascites is collected: collect from mouse peritoneal ascites with No. 9 syringe needles.Collect ascites 2-3 time continuously in a few days, until dead mouse.After the ascites of all collections merged, be distributed into 15ml/ prop up be stored in-20 ℃ standby.
C) ascites purifying: mouse ascites obtains mouse-anti people polyclonal antibody behind reorganization Protein G affinity column (Parmacia company) purifying.Damping fluid is the 0.02M phosphate buffer during affinity chromatography, pH7.0; Eluent is 0.1M glycocoll-hydrochloride buffer, and pH 2.7.
2) bag quilt:
6 * 8 removable battens that ELISA Plate adopts Costar company to produce.Is to add each hole of ELISA Plate behind the 20 μ g/ml with polyclonal antibody that step 1) obtains with 0.05M carbonate buffer solution dilution, every hole 100ul, and absorption is spent the night, wash plate with cleaning buffer solution, spend the night with this damping fluid sealing again, dry after the drying, promptly obtain the polyclonal antibody coated elisa plate.By 48 holes/piece with masking foil pack, vacuum sealing.
3. the TN polyclonal antibody for preparing horseradish peroxidase (HRP) mark
1) resists good SATA (N-hydroxy-succinamide-S-acetyl thio ethyl ester) the solution 20 μ l of adding dilution in the grand antibody-solutions, incubated at room 30 minutes at a good TN of 5mg/ml purifying more;
2) add 100 μ l deacetylation solution (i.e. the oxammonium hydrochloride solution that dissolves with the maleic amide acetate buffer solution), incubated at room 2 hours;
3) separate deacetylation SATA derivant (promptly many anti-) and oxammonium hydrochloride with desalting column;
4) with above-mentioned how anti-solution 2ml and 5mg with the HRP of maleic amide activation room temperature reaction 1 hour, promptly obtain HRP enzyme labeling polyclonal antibody.
5) purifying HRP enzyme is marked polyclonal antibody: remove free HRP with the polyacrylamide prepacked column, obtain mole ratio and mark the TN polyclonal antibody near 1: 8 enzyme.
6) assembling: mark the TN polyclonal antibody to suitable working concentration with the enzyme that the damping fluid dilution that contains 10% hyclone is obtained by step 5), press the packing of 6ml/ bottle, be stored in 4 ℃.
4. preparation quality-control product:
Collect healthy normal human serum (being negative patient), in-20 ℃ of preservations through mensuration such as HIV, liver viroid, AIDS, syphilis.Each serum of accumulation is mixed, measure TN concentration, and the concentration of two parts of pooled serums is transferred to 10 ± 2ng/ml, 50 ± 10ng/Lml scope, be prepared into low, high quality-control product with the PBS damping fluid that contains 3%BSA with the ELISA method.Requirement packing by each kit is stored in 4 ℃ after the freeze drying.Contain each one bottle of low concentration quality-control product and high concentration quality-control product in each kit.
5. preparation substrate solution: 3% superoxol of phosphoric acid-citrate buffer solution (pH5.0) preparation, press the packing of 3ml/ bottle.
6. preparation colour developing liquid: TMB (0.1mg/ml) methanol solution is pressed the packing of 3ml/ bottle.
7. preparation reaction terminating liquid: 2M H
2SO
4, press the packing of 3ml/ bottle.
8. 1% polysorbas20 solution of preparation cleaning buffer solution (20 times of concentrates): PBS (pH7.4) preparation is pressed the packing of 15ml/ bottle.
Use the quality testing that the technology of the present invention prepares the inspection-free test agent box of the quantitative enzyme of TN:
1) accuracy: be divided into three parts of pooled serum samples, every part of 1ml after getting three parts of testing samples mixing.Add 0,20 respectively, the pure product of TN of 100ng, make the recovery test serum specimen
#1,
#2,
#3.The by specification operation steps is measured and result of calculation.Then by recovery computing formula calculate recovery rate.Sample
#2,
#3 the recovery is respectively 96.4% and 98.7%, and average recovery rate is 97.5%, and promptly the proportional jitter of kit is 2.5%, and accuracy is 97.5%.
2) precision: randomly draw 20 box different batches kits, use with a sample to be tested by specification operation steps and carry out replication.Calculate each measurement result, obtain average, SD and coefficient of variation CV.CV≤15% between the Precision test result demonstration is criticized
3) range of linearity: the pure product solution that is diluted to a series of variable concentrations with the pure product of TN: 300ng, 150ng, 75ng, 37.5ng, 18.8ng, 9.5ng, 4.8ng, 2.4ng, 1.2ng.Operation steps is measured to specifications.With concentration is that horizontal ordinate, absorbance are the ordinate curve plotting.The highest detection higher limit is 150ng/ml in the range of linearity, and the lowest detection lower limit is 2.4ng/ml.The range of linearity of kit is 2.4～75ng/ml.
4) detection sensitivity: according to above-mentioned range of linearity measurement result, the detection sensitivity of this kit is 2.4ng/ml.
5) specificity: be divided into four parts of pooled serum samples, every part of 1ml after getting four parts of testing samples mixing.Make the interference test serum specimen after adding tissue-type plasminogen activator (tPA), plasmin (Plasmin) or the fibronectin (FN) of 50ng dosage respectively
#1,
#2,
#3, do not add any chaff interference
#4 pooled serum samples are as basic sample.The by specification operation steps is measured and result of calculation.Calculate jamming rate by the interference test computing formula then.Sample
#1,
#2,
#3 mushing error is all less than 1.5%.
Embodiment two: the use-case of the inspection-free test agent box of the quantitative enzyme of TN
1. cleaning buffer solution preparation: 20 times of dilutions of concentrated cleaning buffer solution adding distil water that kit is provided.
2. first freeze-dried powder of the standard items that kit provided (concentration 75ng/ml) dissolve with 500 μ l cleaning buffer solutions, carry out doubling dilution then 5 times.Typical curve in the kit is made up of the TN standard items of 6 variable concentrations, and typical curve each point concentration is respectively 75ng/ml, 37.5ng/ml, 18.8ng/ml, 9.5ng/ml, 4.8ng/ml, 2.4ng/ml.
3. the dilution of quality-control product: low, the high quality-control product freeze-dried powder that kit is provided dissolves with 110 μ l cleaning buffer solutions respectively.
4. antigen-antibody reaction: in the micropore of the antibody sandwich plate that kit provides, add standard items, quality-control product that 100 μ l have diluted good variable concentrations respectively, or the test serum sample, 37 ℃ of water bath heat preservations 60 minutes.Repeat to wash plate 4 times with cleaning buffer solution then, each 3min.
5. integrated enzyme reaction: HRP-polyclonal antibody solution is added each hole, every hole 100 μ l, 37 ℃ of water bath heat preservations 60 minutes.Repeat to wash plate operation 4 times, each 3min.
6. chromogenic reaction: every hole adds substrate solution, each 50 μ l of TMB colour developing liquid successively, 37 ℃ of water bath heat preservations 10 minutes, and every hole adds 50 μ l reaction terminating liquids again and finishes reaction.
7. colorimetric:, measure OD mean value at 450nm with microplate reader with the light absorption value zeroing in blank hole; Write down each hole light absorption value; Calculate the mean value of diplopore standard items OD value.
8. the result calculates:
1) drafting of typical curve: with standard items concentration is that the average OD value that horizontal ordinate, standard items are measured is an ordinate, draws the typical curve of this mensuration; Basis of calculation curvilinear regression coefficients R
2, work as R
2This experiment in＞0.98 o'clock effectively.
2) judge of quality-control product concentration:, read corresponding quality-control product concentration value from typical curve according to the OD value of quality-control product.When low value quality-control product concentration when 10 ± 2ng/ml, high value quality-control product concentration are in 50 ± 10ng/ml scope, this mensuration is judged to effectively;
3) calculate the test serum sample concentration: when typical curve and quality-control product all are determined when effective, calculate TN concentration the test serum sample from typical curve according to the OD value of sample to be tested.
Claims (9)
1. enzyme-linked immunologic detecting kit that is used to diagnose early ovarian cancer is characterized in that it is made up of by TN polyclonal antibody, quality-control product and the auxiliary reagent of plate, horseradish peroxidase-labeled a kind of ovarian cancer protein mark TN standard items, polyclonal antibody bag.
2. according to the kit of claim 1, wherein said TN standard items are pure product preparations of TN of obtaining through saturated ammonium sulphate, ion-exchange chromatography, the separation and purification of lysine affinity chromatography with human plasma.
3. according to the kit of claim 2, wherein said polyclonal antibody bag is made by the TN polyclonal antibody coated elisa plate that plate obtains with the pure product immune animal of TN.
4. according to the kit of claim 3, purifying obtains the ascites that wherein said TN polyclonal antibody obtains from the pure product immune mouse of TN.
5. according to the kit of claim 4, wherein said polyclonal antibody bag is that the TN polyclonal antibody is splashed in each hole of ELISA Plate after with the carbonate buffer solution dilution by the making of plate, through absorption, wash plate, seal, dry up etc. and make after the step process.
6. according to the kit of claim 5, the TN polyclonal antibody of wherein said horseradish peroxidase-labeled is to use the TN polyclonal antibody, prepares through horseradish peroxidase-labeled.
7. according to the kit of one of claim 1-6, wherein said quality-control product obtains through the dilution allotment with the PHS.
8. according to the kit of claim 7, wherein said auxiliary reagent comprises: integrated enzyme reaction substrate solution, colour developing liquid, reaction terminating liquid and cleaning buffer solution.
9. be used to detect and/or diagnose the purposes of early ovarian cancer according to the kit of one of claim 1-8.
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|---|---|---|---|
| CN 03136488 CN1266475C (en) | 2003-06-05 | 2003-06-05 | Four mucin enzyme linked immunosorbent assay reagent box for early diagnosing oophoroma |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03136488 CN1266475C (en) | 2003-06-05 | 2003-06-05 | Four mucin enzyme linked immunosorbent assay reagent box for early diagnosing oophoroma |
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| Publication Number | Publication Date |
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| CN1553190A true CN1553190A (en) | 2004-12-08 |
| CN1266475C CN1266475C (en) | 2006-07-26 |
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|---|---|---|---|
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102707068A (en) * | 2012-05-31 | 2012-10-03 | 北京大学 | Application of complement factor H (CFH) to genetic expression products of methamphetamine (METH) addiction patient |
| CN103983791A (en) * | 2014-05-29 | 2014-08-13 | 深圳市柏明胜医疗器械有限公司 | Human CA125 protein quantum dot labeled double-sandwiched immunoassay detection kit |
| CN106124765A (en) * | 2006-01-04 | 2016-11-16 | 富士瑞必欧美国公司 | The application in ovarian cancer is assessed of HE4 and other biochemical marker |
| CN111735949A (en) * | 2020-07-17 | 2020-10-02 | 北京信诺卫康科技有限公司 | Wnt7a and CA125 combined used as early ovarian cancer biomarker and kit |
| CN111961723A (en) * | 2020-07-17 | 2020-11-20 | 潘志文 | Tumor marker for noninvasive detection of early ovarian cancer diagnosis and kit |
| CN113325178A (en) * | 2021-05-27 | 2021-08-31 | 江苏省肿瘤医院 | Detection kit for early diagnosis and prognosis evaluation of ovarian cancer |
-
2003
- 2003-06-05 CN CN 03136488 patent/CN1266475C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106124765A (en) * | 2006-01-04 | 2016-11-16 | 富士瑞必欧美国公司 | The application in ovarian cancer is assessed of HE4 and other biochemical marker |
| CN102707068A (en) * | 2012-05-31 | 2012-10-03 | 北京大学 | Application of complement factor H (CFH) to genetic expression products of methamphetamine (METH) addiction patient |
| CN102707068B (en) * | 2012-05-31 | 2015-03-18 | 北京大学 | Application of complement factor H (CFH) to genetic expression products of methamphetamine (METH) addiction patient |
| CN103983791A (en) * | 2014-05-29 | 2014-08-13 | 深圳市柏明胜医疗器械有限公司 | Human CA125 protein quantum dot labeled double-sandwiched immunoassay detection kit |
| CN111735949A (en) * | 2020-07-17 | 2020-10-02 | 北京信诺卫康科技有限公司 | Wnt7a and CA125 combined used as early ovarian cancer biomarker and kit |
| CN111961723A (en) * | 2020-07-17 | 2020-11-20 | 潘志文 | Tumor marker for noninvasive detection of early ovarian cancer diagnosis and kit |
| CN113325178A (en) * | 2021-05-27 | 2021-08-31 | 江苏省肿瘤医院 | Detection kit for early diagnosis and prognosis evaluation of ovarian cancer |
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| Publication number | Publication date |
|---|---|
| CN1266475C (en) | 2006-07-26 |
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