CN103163304A - Method for rapidly detecting follicle stimulating hormone (FSH) in urine sample in a quantitative mode - Google Patents
Method for rapidly detecting follicle stimulating hormone (FSH) in urine sample in a quantitative mode Download PDFInfo
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- CN103163304A CN103163304A CN2011104271738A CN201110427173A CN103163304A CN 103163304 A CN103163304 A CN 103163304A CN 2011104271738 A CN2011104271738 A CN 2011104271738A CN 201110427173 A CN201110427173 A CN 201110427173A CN 103163304 A CN103163304 A CN 103163304A
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Abstract
The invention discloses a method for rapidly detecting follicle stimulating hormone (FSH) in a urine sample in a quantitative mode. The method includes that FSH content is firstly detected, and then a displayed number value range displayed on a reagent card is read by combining a standard curve or a color chart in an immune chromatography interpretoscope. The judging method is easy and convenient to operate and reasonable in design, a patient can detect the FSH content of the patient anywhere at any time to know pituitary endocrine functions and also can indirectly know a hypothalamus and ovarian functional state, the method for rapidly detecting the FSH in the urine sample in the quantitative mode has many helps for pituitary or hypothalamic amenorrhea differential diagnosis.
Description
Technical field
The invention belongs to the technical field of medicine equipment external diagnosis reagent, specifically relate to the method for FSH (FSH) in a kind of Quantitative detection urine sample.
Background technology
FSH (FSH) is by a kind of hormone of anterior pituitary basicyte secretion, and composition is glycoprotein.Main Function is for promoting follicle maturity.FSH regulates and controls the relevant a series of physiology courses of growth, growth, sexal maturity in puberty and reproduction of human body, particularly stimulates the maturation of reproduction cell.FSH (FSH) promotes whole ovary to grow up by promoting stratum granulosum of ovarian follicle hyperplasia differentiation.Acting on convoluted tubule of testis can promote sperm to form.Injection FSH only increases Follicle number, and follicle maturity be there is no effect.The follicle stimulating hormone releasing hormone of hypothalamus secretion is controlled the secretion of follicular stimulating hormone.In the menstrual cycle, in blood, FSH concentration and every day are changed with the cycle by the amount of the FSH of homaluria.After menelipsis, in blood and urine, the FSH discharge rate increases.
Promote the production of sperm pipe to form and Spermatogenic action the male sex, its function is to promote the maturation of convoluted tubule of testis and the generation of sperm; In the women, the function of FSH is to promote Follicle development and maturation, and collaborative interstitialcellstimulating hormone (ICSH) (LH) impels fully-developed ovarian follicle secretion estrogen and ovulation, the formation of participation menorrhea.Its generation is subjected to the control of hypothalamus gonadotropin releasing hormone, is subjected to simultaneously the feedback regulation of ovary female hormone (E2).FSH plays a decisive role to property, the reproductive function of men and women's both sexes.The concentration of blood FSH the preovulatory phase be 1.5~10mIU/ml, be 8~20mIU/ml the onset of ovulation, the ovulation later stage is 2~10mIU/ml.Generally with 5~40mIU/ml as normal value.
By FSH is measured, can understand pituitary endocrine function, also can indirectly understand the functional status of hypothalamus and ovary.This antidiastole to hypophysis or the amenorrhoea of hypothalamus type is helpful.
FSH detects abnormal clinical meaning
The FSH level increases and sees
A) male testical seminoma;
B) prompting women ovarian function height is low, and is as fat in congenital anovaria or ovarian hypoplasia (as Turner syndrome etc.), primary amenorrhea, primary hypogonadism, nervous centralis and hypophysis sex premature, ovary (sexual hypofunction) etc.;
C) menopausal syndrome or menopausal women.
The reduction of FSH level sees
A) the prompting pathology may be returned hypothalamus at hypophysis, urgees hypogonadism etc. as sheehan's syndrome, pituitary chromophobe cell knurl, basicyte knurl (Cushing syndrome), oxyphilic granular cell adenoma's (acromegalia) and primary hypophysis;
B) dysdrophia adiposogenitalis, hypothalamic lesion (as galactorrhea-amenorrhea syndrome, PCOS) etc.
C) long-term taking contraceptive is widely applied sex hormone.
Immunochromatography reagent due to it easy and simple to handle, have directly perceived, fast, the advantages such as detection efficiency is high, method is easy, pollution-free, expense is cheap, highly sensitive, high specificity, be widely used in clinical diagnosis, field condition and familial self and detect.
The reagent that immunochromatography detects FSH uses double antibody sandwich method.After test fluid was soaked or splashed into the sample pad of test strips, by the capillary action of water-absorption fiber, test fluid is drenched gold-marking binding pad rapidly, and the collaurum bond on gold-marking binding pad is dissolved, and moves forward along chromatographic material with test fluid.If there is determined antigen Ag in test fluid, it will with the collaurum bond in antibody generation specific immune response, form the immune complex of collaurum bond-determined antigen.Then flow through with test fluid and be fixed on the capture agent of T line on the Nc film.The capture agent of T line is caught collaurum bond-determined antigen immune complex, namely with this immune complex generation specific immune response, form the immune complex of collaurum bond-determined antigen-capture agent, thereby make collaurum bond-determined antigen immune complex be trapped in T line place.Test fluid continues reach, when being fixed with the C line of sheep anti mouse I gG antibody on the Nc film of flowing through, sheep anti-mouse antibody generation immune response on the collaurum bond that does not react with determined antigen Ag and C line, thus make free collaurum bond be trapped in C line place.This moment, T line and C line place all had the collaurum bond to be detained, and namely T line and C line place all can observe color (redness or purple), represent the positive result of testing result, namely in test fluid, determined antigen are arranged; If only C line place observes color, and T line place does not observe color, represents the negative result of testing result, namely in test fluid without determined antigen; In liquid to be measured, FSH content is different, and the T line color depth that produces is not identical yet, and along with the increase of FSH content, the T line color will be darker.Yet, can cause high-caliber " hook effect (hook effect) " at some sample middle and high concentration FSH, be that FSH concentration is when being far longer than the upper limit of detection of colloidal gold strip, FSH records result and descends with the increase of concentration on the contrary, the T line color shoals on the contrary, this just causes the low-level illusion of FSH in clinical diagnosis, makes the diagnosis to patient beyond thought problem occur.After this patient's urine dilution, can find that its HCG level reality is very high.The erroneous judgement, the false negative result that produce for overcoming hook effect, the sample dilution becomes a kind of conventional means of this area.
Summary of the invention
The object of the present invention is to provide the method for FSH (FSH) in a kind of Quantitative detection urine sample.
Realize that above-mentioned purpose technical scheme of the present invention is, a kind of method of FSH (FSH) in Quantitative detection urine sample, at first described method measures FSH content, the numerical range that then shows on the typical curve in binding immunoassay chromatography interpretoscope or colorimetric card reading reagent card.
Wherein, the mensuration of FSH content is to measure by methods known in the art, comprises with FSH reagent card, reagent strip or reagent pen measuring.
Described colorimetric card is drawn by following method: take National reference as reference, it is 0mIU/ml, 5mIU/ml, 10mIU/ml, 25mIU/ml, 40mIU/ml that the FSH standard solution is diluted to concentration with normal person's urine (feminine gender) or cow's serum, balance is to room temperature, the reagent strip for preparing is kept flat, add standard items to be checked, static 10 minutes, the color that shows according to testing result was depicted as the FSH colorimetric card
simultaneously, typical curve in the immunochromatography interpretoscope is drawn by following method: take National reference as reference, it is 0mIU/ml that the FSH standard solution is diluted to concentration with normal person's urine (feminine gender) or cow's serum, 5mIU/ml, 10mIU/ml, 15mIU/ml, 20mIU/ml, 30mI U/ml, 40mIU/ml series standard product, balance is to room temperature, the reagent strip for preparing is kept flat, add standard items to be checked, put into supporting immunochromatography interpretoscope after static 10 minutes and carry out reading, with the GOD value (X) that records as horizontal ordinate, with FSH content (y) as ordinate, draw the typical curve of instrument internal memory.
Wherein, the mensuration of FSH content can by comparing with colorimetric card, draw content; Or by the reading in the immunochromatography interpretoscope, the horizontal ordinate on the typical curve in this reading and immunochromatography interpretoscope is complementary, the ordinate reading on typical curve is exactly the content of FSH.
The present invention compared with prior art, this determination methods is easy and simple to handle, reasonable in design, the patient can be at any time, everywhere its FSH content is detected, to understand pituitary endocrine function, also can indirectly understand the functional status of hypothalamus and ovary, this antidiastole to hypophysis or the amenorrhoea of hypothalamus type is helpful.
Description of drawings
Fig. 1 is colorimetric card.
Embodiment
For ease of the understanding of technical solution of the present invention, be introduced below in conjunction with concrete embodiment.
The FSH reagent strip comprises loading pad, note thing pad, cellulose nitrate (NC) film, inhales sample pad and liner plate, the mouse-anti FSH-β monoclonal antibody of coated colloid gold label on labeling pad, be coated with respectively the detection line of mouse-anti FSH-Alpha antibodies formation and the matter line processed that sheep anti-mouse igg antibody consists of on the NC film simultaneously.
Described reagent strip is prepared by following method:
(1) preparation of colloid gold label mouse-anti FSH-β labeling of monoclonal antibody thing pad (2)
prepare with gold chloride-trisodium citrate reduction method the colloidal gold solution that diameter is 40nm, after completing, preparation gets three parts of collaurums, pH value of solution can be transferred to pH7.5 with 0.2MK2CO3 respectively, pH8.5, pH9.5, then solution is placed on magnetic stirring apparatus and slowly stirs, by every 100ml solution 0.5mg, 1.0mg, 1.5mg mouse-anti FSH-β monoclonal antibody slowly is added drop-wise in colloidal gold solution, continue to stir 1 hour, adding final concentration is 0.2%BSA again, 0.1% PEG 20000 sealed 30 minutes, 12000 left the heart 30 minutes, abandon supernatant, redissolve to 76.5ml with the collaurum working fluid, ratio in 1ml solution paving 16cm2 is layered on nonwoven fabrics equably, put again 20~25 ℃ of temperature, humidity was less than 30% drying room drying 2~4 hours, make labeling pad, standby.
(2) mouse-anti FSH-Alpha antibodies is coated
With 0.01M pH7.2PBS, mouse-anti FSH-Alpha antibodies is diluted to respectively 1.0mg/ml, 1.5mg/ml, 2.0mg/ml, then with spray film instrument, mouse-anti FSH-Alpha antibodies coating buffer is pressed the coated T line (6) that forms of 1.3ul/cm line on the NC film, coated sheep anti mouse I gG antibody forms C line (7) simultaneously, after coated completing, the NC film was placed in the drying room drying 3~4 hours, standby.
(3) assembling of test strips
20~25 ℃ of temperature, humidity is less than getting liner plate (5) in 30% hothouse, and paste at the middle part that coated NC film is placed on plastic support board, in NC film T line one side overlap joint labeling pad, pastes loading pad (1) at labeling pad opposite side overlap joint; Inhale sample pad (4) in NC film C line one side apart from overlapping edges.Then with cutter, the plastic plate that posts is cut into the test strips of proper width.The test strips that cuts also can be packed in plastic clip (pen), forms to detect reagent card (pen).
The drafting of instrument memory standard curve
Take National reference as reference, it is 0mIU/ml, 5mIU/ml, 10mIU/ml, 15mIU/ml, 20mIU/ml, 30mIU/ml, 40mIU/ml series standard product that the FSH standard solution is diluted to concentration with normal person's urine (feminine gender) or cow's serum, balance is to room temperature, the reagent strip for preparing is kept flat, add standard items to be checked, put into supporting immunochromatography interpretoscope after static 10 minutes and carry out reading, to record GOD value (X) as horizontal ordinate, as ordinate, draw instrument memory standard curve with FSH content (y).
Clinical sample detects
Detect 30 parts of clinical samples according to above-mentioned detection method, take the biological FSH quantitative reagent of Beijing North as contrast, result shows that the sensitivity of this product reaches 25mIU/ml, specificity 100%, and the biological FSH quantitative reagent of Beijing North as a result coincidence rate be 98.5%.
The drafting of colorimetric card
The testing result Show Color that concentration is 0mIU/ml, 5mIU/ml, 10mIU/ml, 25mIU/ml, 40mIU/ml is produced colorimetric card.
The result and the colorimetric card that detect reagent are compared, and interpretation goes out the numerical range of testing result.Coordinate simultaneously FSH secretion information slip to judge whether s own situation is abnormal, obtain treatment in order in time seek medical advice.
Technique scheme has only embodied the optimal technical scheme of technical solution of the present invention, and some changes that those skilled in the art may make some part have wherein all embodied principle of the present invention, within belonging to protection scope of the present invention.
Claims (4)
1. the method for FSH (FSH) in a Quantitative detection urine sample, is characterized in that, at first described method measures FSH content, the numerical range that then shows on the typical curve in binding immunoassay chromatography interpretoscope or colorimetric card reading reagent.
2. detection method according to claim 1, is characterized in that, wherein the measurement of FSH content is to measure by FSH reagent (card or bar or pen).
3. detection method according to claim 1, it is characterized in that, described colorimetric card is drawn by following method: take National reference as reference, it is 0mIU/ml, 5mIU/ml, 10mIU/ml, 25mIU/ml, 40mIU/ml that the FSH standard solution is diluted to concentration with normal person's urine (feminine gender) or cow's serum, balance keeps flat to room temperature the reagent for preparing, and adds standard items to be checked, static 10 minutes, the color that shows according to testing result was depicted as the FSH colorimetric card.
4. detection method according to claim 1, it is characterized in that, typical curve in the immunochromatography interpretoscope is drawn by following method: take National reference as reference, it is 0mIU/ml that the FSH standard solution is diluted to concentration with normal person's urine (feminine gender) or cow's serum, 5mIU/ml, 10mIU/ml, 15mIU/ml, 20mIU/ml, 30mIU/ml, 40mIU/ml series standard product, balance is to room temperature, the reagent strip for preparing is kept flat, add standard items to be checked, put into supporting immunochromatography interpretoscope after static 10 minutes and carry out reading, with the GOD value (X) that records as horizontal ordinate, with FSH content (y) as ordinate, draw the typical curve of instrument internal memory.
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Cited By (3)
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TWI570409B (en) * | 2014-01-14 | 2017-02-11 | Tanaka Precious Metal Ind | Immunochromatographic assay, immunochromatographic assay, and immunochromatographic assay kits |
CN111596076A (en) * | 2020-06-04 | 2020-08-28 | 昆明天沃生物科技有限公司 | Method for detecting sheep fertility |
CN111596074A (en) * | 2020-06-04 | 2020-08-28 | 昆明天沃生物科技有限公司 | Method for detecting fertility of cattle |
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
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TWI570409B (en) * | 2014-01-14 | 2017-02-11 | Tanaka Precious Metal Ind | Immunochromatographic assay, immunochromatographic assay, and immunochromatographic assay kits |
TWI585410B (en) * | 2014-01-14 | 2017-06-01 | Tanaka Precious Metal Ind | Immunochromatographic assay, immunochromatographic assay, and immunochromatographic assay kits |
CN111596076A (en) * | 2020-06-04 | 2020-08-28 | 昆明天沃生物科技有限公司 | Method for detecting sheep fertility |
CN111596074A (en) * | 2020-06-04 | 2020-08-28 | 昆明天沃生物科技有限公司 | Method for detecting fertility of cattle |
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Application publication date: 20130619 |