CN101750501A - Protein chip for female infertility detection and kit thereof - Google Patents
Protein chip for female infertility detection and kit thereof Download PDFInfo
- Publication number
- CN101750501A CN101750501A CN200810204669A CN200810204669A CN101750501A CN 101750501 A CN101750501 A CN 101750501A CN 200810204669 A CN200810204669 A CN 200810204669A CN 200810204669 A CN200810204669 A CN 200810204669A CN 101750501 A CN101750501 A CN 101750501A
- Authority
- CN
- China
- Prior art keywords
- detection
- antigen
- protein
- female infertility
- protein chip
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention discloses a dedicated multi-index protein chip for female infertility detection and a kit thereof. The protein chip consists of a substrate, protein detection indexes arranged on the substrate in an array mode and a contrast coating for the detection indexes. The protein detection indexes cover ovary antigen (OAg), endometrium antigen (EAg), sperm antigen (SAg), zona pellucida antigen (ZAg), human chorionic gonadotropin antigen (HCG), trophoblast antigen (TAg) and cardiolipin antigen (CAg). The present invention overcomes the defects of the existing autoimmunological detection products on market, such as single detection function, lagged detection indexes and no integration. Besides the indexes of the product (patent application No: 200510102466.3), the protein chip also has the ACA index. The density of all indexes can be quantitatively detected, and the cross reaction of all indexes is eliminated. The present invention not only has high specificity and high sensitivity, but also has high accuracy and precision.
Description
Technical field
The present invention relates to protein chip disease detection product, specifically, relate to a kind of protein chip for female infertility detection and kit thereof.
Background technology
According to World Health Organization (WHO) prediction,, infertilely will become the third-largest disease that is only second to tumour and cardiovascular and cerebrovascular diseases in 21 century.Authority's data presentation, unwillingly the infertile incidence of property is 7%～46% in the married couple.Although different regions there are differences, generally be considered to 8%～15%.Present domestic infertile patient accounts for 10% of married woman's sum, will account for 15% abroad, and the incidence of disease all has ascendant trend year by year.Generally speaking, the infertile factor wife's side will account for 2/3, and the bridegroom's or husband's side will account for 1/3.
Women's infertility has the branch of former and secondary from time sequencing.Former is meant after women's sexal maturity never conceivedly or gave birth to, and secondary is meant that past attempts had gestation or BH and occurred infertile once more.From pathology, the reason of female infertility is concluded organic disease, endocrine factors, immune factor and mental element etc.Article (the immunologic test interpretation of result of the sterile female patient of 760 routine secondary infertilities according to people such as Huang Shaokun, China's aristogenesis and Journal of Heredity, 1006-9534 (2003) 01-0076-02) shows, the female infertility that the immunity factor causes is that most of unknown cause is infertile, secondary infertility is sterile and the main cause of recurrent abortion, and diagnosis and the treatment of the secondary infertility infertile patients being carried out a whole set of immune factor are necessary.
The immunology diagnosis of female infertility is the new diagnostic method of just carrying out in recent years.Mainly comprise AOAb (AOA at present, anti-ovarian antibody), AEA (AEA, anti-endometrial antibody), AsAb (ASA, anti-spermatozoa antibody), anti-vitellary membrane antibody (AZP, anti-zona pellucidantibody), anticardiolipin antibody (ACA, anti-cardiolipin antibody), anti-hCG antibodies (HCG-ab, human chorionic gonadotropin antibody), anti-trophocyte membrane antibody indexs such as (ATA, anti-trophoblast membrane antibody).
The generation of AOAb (AOA) is owing to reasons such as ovary damage, infection, inflammation cause the excessive of ovary antigen to cause.The high expressed that AOAb (AOA) wherein arranged among the infertile and miscarriage patient of about 42%-52%.The expression of AOA can further increase the weight of the damage of ovary, and causes the function of uterus, placenta unsound, thereby causes infertile and miscarriage.
AEA (AEA) is because the uterus under pathological state, as endometritis, endometriosis and uterus adenomyosis etc., can change into antigen or haptens, stimulates body and the antibody that produces.In addition, during induced abortion dilatation and curettage, blastular also may produce antibody as the antigenic stimulus body.In case have AEA to exist in women's body, just can cause infertile, stop pregnant or miscarry.
Sperm is as a kind of antigen of uniqueness, and can cause self or homoimmune reaction after body immune system contacts, and produces AsAb (ASA), infertile 10%～30% of the infertile patient that accounts for due to the AsAb.All may there be AsAb in men and women's body.Be present in serum or the refining at male anti-sperm antibody, then be present in serum or the cervical mucus in the women.ASA and cervical mucus protein combination can disturb sperm to pass the cervical mucus onset of ovulation; The opsonic action of ASA can promote the local phagocyte of genital tract engulfing sperm; The antibody of anti-sperm head can also be blocked smart ovum combination, causes the melting of embryonated egg of complement-mediated, and the infringement embryo implants and pre-embryo is grown and caused early abortion.
Anti-vitellary membrane antibody (AZP) is produced by the reaction of women's autoimmunity.Oolemma is the glycoprotein that is coated in the ovum surface) antibody.Each menstrual cycle always has some ovarian follicles to become atretic follicle, and oolemma wherein is if any activity, can become antigenic stimulus and produces anti-vitellary membrane antibody, or owing to infect and cause the oolemma sex change, stimulate body to produce anti-vitellary membrane antibody.Anti-vitellary membrane antibody has hidden the sperm-receptor that is positioned on the oolemma, makes sperm can not be familiar with ovum, also just has no way of combining with ovum.Simultaneously antibody can stable transparent belt surface structure, thereby can resist the dissolution of acrosin to oolemma, makes sperm not penetrate oolemma.As being fertilized, because of stablizing of oolemma structure, cause the embryo and be locked in the oolemma, and can't implantation.
Anticardiolipin antibody (ACA) is a kind of with the autoantibody of cuorin electronegative on blood platelet and the endothelial cell membrane as target antigen.Be common in systemic loupus erythematosus (SLE) and other autoimmune diseases.This antibody and thrombosis, blood platelet, spontaneous abortion or FDIU are in close relations.ACA positive person stillborn foetus incidence is about 76%, and spontaneous abortion repeatedly and FDIU and ACA are remarkable positive correlation.The pregnant woman of ACA positive rate is in the early stage easily recurrent abortion of gestation, and in gestation, the later stage, intrauterine fetal death easily took place, particularly in the ACA-IgG level to height rising person.Oshiro was to detecting ACA and the women that miscarries more than 2 times carries out retrospective study and finds, intrauterine fetal death takes place the positive women of ACA is the type of sending out early abortion pregnant and have more characteristics.And the research of Lockshin etc. announcement ACA can more early predict spontaneous abortion more sensitively.
Human chorionic gonadotrophin (HCG) is a major hormone of keeping early pregnancy.But the women that history of spontaneous abortion, history of artificial abortion and biochemical pregnant history are arranged, in the miscarriage process, the HCG in the chorionic villi may produce antibody as the antigenic stimulus parent.Once accepted the HCG injection, to promote the women of ovulation, the anti-HCG antibody in the body also might be positive.This type of patient may show as infertile or habitual abortion clinically.In addition, in women's serum of unknown cause miscarriage, anti-trophocyte's antibody (ATAb) increases than normal pregnancies is obvious, this antibody increase and miscarry between close ties are arranged.Its mechanism may be relevant with the minimizing of blocking antibody.
Detection method to the female infertility amynologic index mainly concentrates on immunofluorescence technique or indirect immunofluorescence at present.Complex operation and time-consuming.The ELISA detection kit of external existing part index number detects but can only carry out single index, and cost and time cost are all than higher.Therefore demand urgently a kind of can high flux, rapid and precise detection method.
Biochip is the high-level efficiency detection method that developed recently gets up, and can obtain desired testing result fast, improves detection efficiency greatly, saves cost.Number of patent application is that 200510102466.3 one biochip that can detect 6 infertile indexs has met fast, high flux, high specific and sensitivity market demands.But still have certain defective, as: 1. this kit lack to the detection of present widely used ACA index 2. this kit be qualitative kit, only can judge positive and negative, can not accomplish detection by quantitative.And detection by quantitative for the evaluation of patient disease's order of severity with and the corresponding treatment method be very significant.Therefore, present detection means is still waited to improve.
Summary of the invention
The present invention will provide a kind of multi-index protein chip and kit thereof that female infertility detects that be specifically designed to.Solve single, the detection index hysteresis of autoimmunity testing product measuring ability in the market, and do not have integrated shortcoming.Except adding the ACA index that number of patent application is 200510102466.3 a products shortage, also can accomplish the concentration of each index of detection by quantitative, and eliminated cross reaction between index, and not only accomplished high specific and high sensitivity, also have very high accuracy and degree of accuracy.
Technical scheme of the present invention is as follows: a kind of infertile detection protein chip is provided, be distributed in on-chip Protein Detection index and control test index coating is formed by substrate and array, described Protein Detection index comprises ovary antigen (OvarinAntigen, OAg), endometrium antigen (Endometrial Antigen, EAg), sperm antigen (Spermatozoa Antigen, SAg), oolemma antigen (Zona pellucid Antigen, ZAg), human chorionic gonadotrophin (Human chorionicgonadotropin, HCG), trophocyte's membranous antigen (Trophoblast membrane Antigen, TAg), be fixed on above the substrate with covalent bond or non-covalent bond; Described Protein Detection index also comprise cuorin antigen (CardiolipinAntigen, CAg).
Described substrate is plastics, nylon membrane or nitrocellulose filter.
Above-mentioned control test index coating comprises positive control and negative control; Positive control is a rabbit igg antibody, and negative control is the used dilution of antibody spot sample.
In each array on the above-mentioned coating Protein Detection index each 2-10 repetition arranged, the contrast index 1-5 repetition arranged.
What and number big or small and sample to be checked of array, the substrate size, and the point sample diameter is relevant.
The preference of above-mentioned Protein Detection index and detection arrays is: when Protein Detection index point sample diameter is 0.1mm, and dot spacing 0.6mm, array size is 6mm * 5mm, and 24 arrays can be set on the nitrocellulose filter protein chip.
The present invention also provides the kit that comprises above-mentioned infertile detection protein chip, also comprises standard items and quality-control product; Standard items and quality-control product are bottled, are fixed in the packing box with papery template, plastic formwork or sponge template.
Above-mentioned kit also comprises the sample dilution, enzyme mark working fluid, and concentrated cleaning solution detects liquid; Each liquid is bottled, reinstates papery template, plastic formwork or sponge template with standard items and quality-control product one and is fixed in the packing box.
Each several part is fixed in the packing box by fixed mould, does not limit modes of emplacement, supporting use during detection.
Above-mentioned standard items are made up of the mixed liquid of protein of 5-10 variable concentrations gradient, each mixed liquid of protein container of packing into, and mixed liquid of protein detects indexs by 7 kinds of female infertility marker proteins and mixes by variable concentrations.
Standard items are used for detecting at each sample is doing typical curve after finishing, the protein concentration of the detection index in can the quantitative measurement sample.
Above-mentioned quality-control product is made up of the mixed liquid of protein of 3-6 variable concentrations, each mixed liquid of protein container of packing into, and mixed liquid of protein detects indexs by 7 kinds of female infertility marker proteins and mixes by variable concentrations.
Quality-control product can be monitored the CV value of whole detection system, guarantees the validity of each testing result.
Above-mentioned sample dilution is a kind of protein stabiliser; Can be by 3%BSA and 0.05% antiseptic (Proclin
TM300) phosphate buffer (pH 7.4) is formulated;
Above-mentioned enzyme mark working fluid is by corresponding antigens that contains the HRP mark or monoclonal antibody, and the aforementioned sample dilution of HRP-goat anti-rabbit igg is formed;
Above-mentioned concentrated cleaning solution is the neutral buffered liquid of a kind of PH6.5-PH8.0; It can be phosphate buffer;
Above-mentioned detection liquid refers to matching used two kinds of reagent, a kind of luminol and chemiluminescence intensifier of containing, and another kind contains hydrogen peroxide.
Description of drawings:
Fig. 1 is the outside drawing of protein chip for female infertility detection, and 1 is conversion zone, and the bottom is the substrate of point sample coating.
Fig. 2 is the sample application array synoptic diagram, and the 1st behavior positive control rabbit igg antibody and negative control respectively has 3 to repeat point sample; All the other each row are respectively Protein Detection index point sample OAg, EAg, SAg, ZAg, CAg, HCG and TAg totally 7 marks, two indexs of every row, first row OAg, the EAg, second row SAg, the ZAg, the third line CAg, HCG, fourth line TAg respectively has 5 to repeat point sample.
Fig. 3 is a protein chip for female infertility detection kit synoptic diagram.2 represent concentrated cleaning solution; 3 represent 1 bottle of sample dilution and 1 bottle of enzyme mark working fluid; 4 represent 2 bottles of matching used detectable; 5 represent standard items; 6 represent quality-control product.
Fig. 4 is for detecting the serum synoptic diagram, and what wherein 12 rectangles represented that application of sample zone, place adds is standard items, and each repeats 6 standard items 2 times; What 5 polygons represented that application of sample zone, place adds is quality-control product; What all the other circular expression application of sample zones, place added is sample.
Fig. 5 is the female infertility testing result synoptic diagram that a kind of ACA of being mainly causes.Wherein ACA is positive, and the ASA index is the weak positive, and all the other are negative.
Embodiment:
Be to be understood that, unaccounted normal condition and method among the following embodiment, usually according to the conventional employing method of affiliated field experimenter: as " molecular cloning experiment guide " third edition of Sa nurse Brooker and Russell chief editor, or the step and the condition of advising according to manufacturer.
Embodiment 1:
A kind of preparation of protein chip for female infertility detection
Its step mainly is: (1) prepares the matrix nitrocellulose filter; (2) design chips is determined the arrangement mode and the point sample position of dot matrix; (3) spotting needle from 384 orifice plate sucking-off antibody speckings on nitrocellulose filter; (4) assembling nitrocellulose filter, and seal, dry, packing, preserve.
The above-mentioned methods of making protein chips that is used to detect female infertility mark antibody specifically may further comprise the steps:
1. female infertility marker detection antigen-antibody is definite:
Female infertility mark kit detects determining of antigen:
The antigen that is used to prepare protein chip for female infertility detection all extracts in animal tissue or organ.OAg is from the monkey ovary, and EAg is from the cattle uterus inner membrance, and from the ox ovarian follicle, CAg is from bovine cardiac from people's seminal fluid, ZAg for SAg, HCG from human pregnant urine, TAg from human trophoblastic cells.
Be used for preparing the determining of detection indicator antibody of kit standard items:
The detection antibody that is adopted is right to being the mouse authentic monoclonal antibody, and its accuracy of detection and accuracy obviously are better than polyclonal antibody.
2. point sample prepares the film chip that tool detects the index coating
With Gesim point sample instrument specking, on nitrocellulose filter, latticed form is 10 * 5 rectangular arrays to spotting needle from 384 orifice plate sucking-off antibody speckings, and when spot diameter is 0.1mm, the some horizontal spacing is 0.6mm, and the size of antibody array is 6mm * 5mm; Pad pasting behind the point sample, sealing; At last with chip drying, packing back in 4 ℃ of preservations.
3. protein chip assembling
With having the utensil of separating conversion zone conversion zone is fixed.Assembling mode is seen ZL032298900.Synoptic diagram such as Fig. 1 after assembling is finished.
Embodiment 2:
The preparation of kit
A kind of infertile detection protein chip kit, by infertile detection protein chip, standard items, quality-control product, the sample dilution, enzyme mark working fluid, concentrated cleaning solution detects liquid and forms; Each several part is bottled, is fixed in the packing box (as Fig. 2) with the papery template, does not limit modes of emplacement, supporting use during detection.Below be the preparation method of each several part:
1. infertile detection protein chip is 6 * 8 orifice plates, and as the sheet base, assembling mode is seen ZL032298900 by nitrocellulose filter.Outside drawing is seen Fig. 1 after finishing.
2. standard items provide 6 bottles, every bottle of 0.25ml.
With containing 3%BSA and 0.05% antiseptic (Proclin
TM300) phosphate buffer (pH 7.4) configuration, wherein the final concentration of AOA, AEA, ASA, AZP, ACA, HCG-ab, ATA standard items sees the following form:
3. quality-control product provides 5 bottles, every bottle of 0.2ml.
Quality-control product is divided into 5 concentration, is respectively quality-control product I, quality-control product II, quality-control product III, quality-control product IV, quality-control product V.These five quality-control product concentration level off to respective standard product concentration respectively.
Quality-control product is with containing 3%BSA and 0.05% antiseptic (Proclin
TM300) phosphate buffer (pH 7.4) preparation, following table shows the final concentration and the Quality Control boundary of AOA, AEA, ASA, AZP, ACA, HCG-ab, ATA standard items.
Above-mentioned quality-control product Quality Control boundary assay method is as follows:
Respectively by 2 experimenters according to standard test operation detection quality-control product, repeat 4 times/plate, every day, everyone repeated to survey 2 chip blocks, duplicate detection 10 days is calculated the average and the standard deviation of quality-control product respectively, the Quality Control boundary is
± 15%.
4. sample dilution
By 3%BSA and 0.05% antiseptic (Proclin
TM300) phosphate buffer (pH 7.4) is formulated.Provide 1 bottle, 9ml.Be used for dilute serum sample (volume ratio 1: 1).
5. enzyme is marked working fluid
By 3%BSA and 0.05% antiseptic (Proclin
TM300) phosphate buffer (pH 7.4) wiring solution-forming contains 7 monoclonal antibodies that the female infertility mark is special of HRP mark, and detects antibody HRP-goat anti-rabbit igg.Provide 6ml, 1 bottle.
6. concentrated cleaning solution (10 *)
Be phosphate buffer.Form by following method configuration: potassium dihydrogen phosphate 6.8g, sodium hydrogen phosphate 7.1g, TWEEN-2010ml is dissolved in the 1000ml purified water.It is pure that each chemical substance is analysis.
Provide 50ml, 1 bottle of concentrated cleaning solution.Dilute mixing before using with the 450ml deionized water.
7. detection liquid
Use PIERCE company
ELISA Femto Maximum Sensitivity Substrate is as supporting detection liquid.Each 1 bottle of A liquid, B liquid, every bottle of 1.5ml.
Embodiment 3:
The application of protein chip and kit and detection
The infertile mark antibody in Ag-Ab-ELIAS secondary antibody method detection human blood is mainly adopted in the application that is used to detect infertile protein chip of the present invention.Method is that human serum is added on chip surface, and the antibody in the sample is respectively at the corresponding antigen combination that is fixed on the chip; Dry the back and add the ELIAS secondary antibody reaction, flush away unconjugated two is anti-; Add to detect behind the liquid 1 minute, signal is also collected in scanning, and the signal intensity of sample point is directly proportional with the concentration of antibody in the serum.
In the above-mentioned application that is used for detecting infertile protein chip kit, the concrete using method of its antigen-antibody reaction and detection is as follows:
(1) chip to specking antigen, positive control, negative control seals with 3%BSA solution, room temperature 2 hours, the non-specific site of sealing substrate surface;
(2) behind the drying confining liquid, room temperature was placed 4 hours, carried out drying;
(3) at the conversion zone of chip, add human serum sample through waiting dilute at 1: 20, and 2 repetitions of 6 parts of standard items, 5 parts of quality-control products (add sample loading mode and see synoptic diagram 4); Every hole adds 100ul, places 37 ℃ in isothermal reaction instrument, and oscillating reactions 1 hour is fully reacted antigen and antibody;
(4) dry after, every hole adds ELIAS secondary antibody 100ul, places 37 ℃ in isothermal reaction instrument, oscillating reactions 30 minutes, and vibrate on shaking table with the phosphate buffer of 10 times of dilutions and to wash each 1 minute three times;
(5) with PIERCE company
The matching used detection liquid A liquid of ELISA Femto Maximum Sensitivity Substrate and B liquid mix at 1: 1, and on each conversion zone, every array adds 60ul, carries out chemiluminescence on the adding chip;
(6) carry out chemiluminescent scanning and collect signal with the CCD detector, according to scanning result display analysis testing result.
As Fig. 5 is the female infertility testing result synoptic diagram that a kind of ACA of being mainly causes.
Claims (9)
1. protein chip for female infertility detection, be distributed in on-chip Protein Detection index and control test index coating is formed by substrate and array, described Protein Detection index comprises ovary antigen (Ovarin Antigen, OAg), endometrium antigen ((Endometrial Antigen, EAg), sperm antigen (Spermatozoa Antigen, SAg), oolemma antigen (Zona pellucid Antigen, ZAg), human chorionic gonadotrophin (Human chorionic gonadotropin, HCG), trophocyte's membranous antigen (Trophoblast membrane Antigen, TAg), be fixed on above the substrate with covalent bond or non-covalent bond; It is characterized in that, described Protein Detection index also comprise cuorin antigen (CardiolipinAntigen, CAg).
2. protein chip for female infertility detection as claimed in claim 1 is characterized in that, described substrate is plastics, nylon membrane or nitrocellulose filter.
3. protein chip for female infertility detection as claimed in claim 1 or 2 is characterized in that, described contrast index comprises 1 positive control and 1 negative control; Positive control is a rabbit igg antibody, and negative control is the used dilution of antigen point sample.
4. protein chip for female infertility detection as claimed in claim 3 is characterized in that, each detects index on the described coating 2-10 repetition, and the contrast index has 1-5 repetition.
5. a protein chip for female infertility detection kit is characterized in that, comprises the described protein chip for female infertility detection of claim 1, standard items and quality-control product; Standard items and quality-control product are bottled, are fixed in the packing box with papery template, plastic formwork or sponge template.
6. protein chip for female infertility detection kit as claimed in claim 5 is characterized in that, also comprises the sample dilution, enzyme mark working fluid, concentrated cleaning solution and detection liquid; Each liquid is bottled, is fixed in the packing box with papery template, plastic formwork or sponge template.
7. as claim 5 or 6 described protein chip for female infertility detection kits, it is characterized in that, described standard items are made up of the mixed liquid of protein of 5-10 variable concentrations gradient, each mixed liquid of protein container of packing into, mixed liquid of protein detects indexs by 7 female infertility marker proteins and mixes by variable concentrations.
8. as claim 5 or 6 described protein chip for female infertility detection kits, it is characterized in that, described quality-control product is made up of the mixed liquid of protein of 3-6 variable concentrations, each mixed liquid of protein container of packing into, mixed liquid of protein detects indexs by 7 female infertility marker proteins and mixes by variable concentrations.
9. protein chip for female infertility detection kit as claimed in claim 6 is characterized in that the each several part compound method is as follows:
Described sample dilution is a protein stabiliser;
Described enzyme mark working fluid is by the monoclonal antibody of the corresponding antigens that contains the HRP mark, and the aforementioned sample dilution of HRP-goat anti-rabbit igg is formed;
Described concentrated cleaning solution is the neutral buffered liquid of PH6.5-PH8.0;
Described detection liquid refers to matching used two kinds of reagent, a kind of luminol and chemiluminescence intensifier of containing, and another kind contains hydrogen peroxide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810204669A CN101750501A (en) | 2008-12-16 | 2008-12-16 | Protein chip for female infertility detection and kit thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810204669A CN101750501A (en) | 2008-12-16 | 2008-12-16 | Protein chip for female infertility detection and kit thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101750501A true CN101750501A (en) | 2010-06-23 |
Family
ID=42477791
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200810204669A Pending CN101750501A (en) | 2008-12-16 | 2008-12-16 | Protein chip for female infertility detection and kit thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101750501A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104977400A (en) * | 2015-07-27 | 2015-10-14 | 三明市和众生物技术有限公司 | Infertility joint detection kit and detection method thereof |
| CN105256002A (en) * | 2015-08-31 | 2016-01-20 | 成都中医药大学 | Screening kit for female yang deficiency constitution infertility |
| CN105928928A (en) * | 2016-06-30 | 2016-09-07 | 深圳市亚辉龙生物科技股份有限公司 | Chemiluminescence immune detection kit of anti-trophoblast cell membrane antibody and preparation method of chemiluminescence immune detection kit |
| CN106932572A (en) * | 2017-04-26 | 2017-07-07 | 深圳市博卡生物技术有限公司 | For the kit and its detection method of infertile antibody test |
| CN107561266A (en) * | 2016-06-30 | 2018-01-09 | 深圳市亚辉龙生物科技股份有限公司 | AOAb chemiluminescence immune detection reagent kit and preparation method thereof |
| CN107561293A (en) * | 2016-06-30 | 2018-01-09 | 深圳市亚辉龙生物科技股份有限公司 | Anti-hCG antibodies chemiluminescence immune detection reagent kit and preparation method thereof |
| CN108148758A (en) * | 2016-12-05 | 2018-06-12 | 中国科学院大连化学物理研究所 | A kind of external model method for building up of Extra-villous trophoblasts nano particle exposure |
| CN111280162A (en) * | 2019-08-01 | 2020-06-16 | 常州市妇幼保健院 | Storage method for single sperm or very small amount of sperm and chip carrier |
-
2008
- 2008-12-16 CN CN200810204669A patent/CN101750501A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104977400A (en) * | 2015-07-27 | 2015-10-14 | 三明市和众生物技术有限公司 | Infertility joint detection kit and detection method thereof |
| CN105256002A (en) * | 2015-08-31 | 2016-01-20 | 成都中医药大学 | Screening kit for female yang deficiency constitution infertility |
| CN105256002B (en) * | 2015-08-31 | 2019-01-04 | 成都中医药大学 | A kind of kit for screening of women deficiency of yang matter infertility |
| CN105928928A (en) * | 2016-06-30 | 2016-09-07 | 深圳市亚辉龙生物科技股份有限公司 | Chemiluminescence immune detection kit of anti-trophoblast cell membrane antibody and preparation method of chemiluminescence immune detection kit |
| CN107561266A (en) * | 2016-06-30 | 2018-01-09 | 深圳市亚辉龙生物科技股份有限公司 | AOAb chemiluminescence immune detection reagent kit and preparation method thereof |
| CN107561293A (en) * | 2016-06-30 | 2018-01-09 | 深圳市亚辉龙生物科技股份有限公司 | Anti-hCG antibodies chemiluminescence immune detection reagent kit and preparation method thereof |
| CN108148758A (en) * | 2016-12-05 | 2018-06-12 | 中国科学院大连化学物理研究所 | A kind of external model method for building up of Extra-villous trophoblasts nano particle exposure |
| CN108148758B (en) * | 2016-12-05 | 2021-09-17 | 中国科学院大连化学物理研究所 | In-vitro model establishment method for extravillous trophoblast nanoparticle exposure |
| CN106932572A (en) * | 2017-04-26 | 2017-07-07 | 深圳市博卡生物技术有限公司 | For the kit and its detection method of infertile antibody test |
| CN111280162A (en) * | 2019-08-01 | 2020-06-16 | 常州市妇幼保健院 | Storage method for single sperm or very small amount of sperm and chip carrier |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101750501A (en) | Protein chip for female infertility detection and kit thereof | |
| US7989217B2 (en) | Method for determining hCG levels in fluid samples | |
| Kovalevskaya et al. | Early pregnancy human chorionic gonadotropin (hCG) isoforms measured by an immunometric assay for choriocarcinoma-like hCG | |
| US4952517A (en) | Positive step immunoassay | |
| US20030119050A1 (en) | Flow cytometer for analysis of general diagnostic factors in cells and body fluids | |
| US20160282343A1 (en) | Quantitative lateral flow assay strips for quantitative analysis of an analyte, kits containing such strips and methods of manufacture and use of same | |
| US4874693A (en) | Method for assessing placental dysfunction | |
| EA016348B1 (en) | Assay for determining implantation potential of embryos and method for assisted fertilisation | |
| US20140024057A1 (en) | Systems and methods for detecting animal pregnancy | |
| Khosravi et al. | Immunofluorometry of choriogonadotropin by time-resolved fluorescence spectroscopy, with a new europium chelate as label. | |
| CN101769926A (en) | Integrated test reaction plate of five indicators of prenatal and postnatal care and kit | |
| Kadem et al. | Unusual false-negative serum human chorionic gonadotropin detected by qualitative immunoassay: A case report of two Iraqi women | |
| AU674327B2 (en) | Diagnostic method for determining the susceptibility to delivery and reagent kit for use therefor | |
| Walker et al. | Occult biochemical pregnancy: fact or fiction? | |
| CN103344774A (en) | Chemiluminescence immune assay kit and method for magnetic particle of human estradiol (E2) | |
| Chen et al. | Quantitative analysis of total β-subunit of human chorionic gonadotropin concentration in urine by immunomagnetic reduction to assist in the diagnosis of ectopic pregnancy | |
| CN101871942A (en) | Premature rupture of membrane (PROM) detection kit using ICAM-1 as examination index and preparation method | |
| US6025149A (en) | Urinary screening for down syndrome and other aneuploidies | |
| CN106405117A (en) | Plate type magnetic microparticle chemiluminescence analysis kit for detecting leptin antigen | |
| Ramsay et al. | Urinary steroid evaluations to monitor ovarian function in exotic ungulates: X. Pregnancy diagnosis in Perissodactyla | |
| CN109212195A (en) | A kind of kit quickly detecting people AMH using Magnetism particulate immuno chemistry luminescence method | |
| Vankrieken et al. | Rapid, automated quantification of total human chorionic gonadotropin in serum by a chemiluminescent enzyme immunometric assay | |
| CN101858917A (en) | Kit for detecting premature rupture of membrane by taking Axl as detection index and preparation method thereof | |
| CN1085339C (en) | Contraceptive test paper for safe period and its test method | |
| CN103293310B (en) | A kind of kit detecting premature rupture of fetal membranes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20100623 |