CN201011517Y

CN201011517Y - FSH and LH associated detecting test paper

Info

Publication number: CN201011517Y
Application number: CNU200720005148XU
Authority: CN
Inventors: 万志静; 万志强
Original assignee: WHPM Bioresearch and Technology Co Ltd
Current assignee: WHPM Bioresearch and Technology Co Ltd
Priority date: 2007-02-14
Filing date: 2007-02-14
Publication date: 2008-01-23
Anticipated expiration: 2017-02-14

Abstract

The utility model belonging to the detecting field of collaurum immunity, discloses a FSH and LH joint detecting test paper, which externally detects the FSH and LH levels of urine, a nature determining gynaecological diseases and female ovarian function by the means of collaurum double antibody clamp antigen. The utility model consists of a base plate, a water absorbing plate, a nitrocellulose membrane, an anti FSH trastuzumab Chin Piao pad, an anti LH trastuzumab Chin Piao pad, a sampled liquid sucking layer and a MAX line. The middle part of the base plate is the nitrocellulose membrane, which has a testing line of the FSH trastuzumab, a testing line of the LH trastuzumab and a controlling line of a Goat Anti Mouse polyclonal antibody. One end of the base plate is the water absorbing plate while the other end is the sample liquid sucking layer. The two ends of the nitrocellulose membrane are respectively connected in superposition with the anti FSH trastuzumab Chin Piao pad and anti LH trastuzumab Chin Piao pad. The Chin Piao pads are pressed with the sample liquid sucking layer.

Description

A kind of FSH, LH joint-detection test paper

Technical field

The present invention relates to a kind of FSH, LH joint-detection test paper, belong to the colloid gold immune detection range.

Background technology

Follicular stimulating hormone (FSH) and two kinds of glycoprotein hormoneses of luteotropic hormone (LH) by anterior pituitary produces play an important role on gonad axis, and the women in the childbearing age cyclical variation occurs with the menstrual cycle.FSH acts on the acceptor on the follicular cell, stimulates ovarian follicular growth, growth, maturation, and promotes estrogen secretion.FSH keeps reduced levels in early days at ovarian follicle, with follicular development to late period, estrogen level raises, and FSH slightly descended subsequently, low value occurred to 24 hours onsets of ovulation, raise rapidly immediately, descend again after 24 hours, LH and FSH acting in conjunction cause ovulation, keep low-level luteal phase, and promote female, progestational hormone to synthesize.LH is in low-level in early days at ovarian follicle, rise gradually later on, and occurred the peak simultaneously with FSH in preceding about 24 hours to ovulating, and be the cliffy summit higher than FSH, mxm. rapid drawdown after 24 hours, the corpus luteum later stage descends gradually.Detect and reach FSH, LH level in the urine in the blood, be used for assisting to judge amenorrhoea reason, diagnosis polycystic ovary syndrome and premature ovarian failure, ovarian function deficiency etc. clinically.

Assay method commonly used has biochemical method (as gas chromatographic method method, spectrophotometric method, fluorescence spectrophotometry) and applied immunology method to measure, and mainly is radioimmunoassay (RIA) and enzyme immunoassay (ELISA).These methods need special safeguard, harsh condition, expensive instrument and well-trained personnel, and during operating cost, the sample storage difficulty, be not easy to apply.

Immunoassay provides a new analyzing and testing approach for the detection of FSH, LH, and existing having utilizes the immune colloid gold method to make the report of test paper: 00222540.9 1 kinds of eugenic test papers of Chinese patent; 00112780.2 1 kinds of eugenic test papers of Chinese patent and detection method thereof; 00222539.5 1 kinds of infertilities of Chinese patent detect test paper.Wanhuapuman Bioengineering Co Ltd in nineteen ninety-five just obtained the to ovulate traditional Chinese medicines word authentication code (95) of product defend the accurate word of medicine (Jing Wanhua) S-02 number, and, therefore above-mentioned patent is not constituted infringement with launch products.The difference of the present invention and above-mentioned technology is, is coated with three lines on the nitrocellulose filter, a LH monoclonal antibody test wire,, a FSH monoclonal antibody test wire, article one, control line can detect FSH and LH level in the urine simultaneously, is used to diagnose gynecological disease and ovarian function.

Summary of the invention

The present invention has overcome some problems that exist in the prior art, and a kind of test paper by FSH, LH in the collaurum double antibody folder antigen method detection urine is provided, can external etiologic diagnosis gynecological disease and women's ovarian function.

For solving the problems of the technologies described above, the present invention is achieved by the following technical solutions:

FSH, LH joint-detection test paper base plate middle part are nitrocellulose filter, an anti-FSH monoclonal antibody test wire, an anti-LH monoclonal antibody test wire and a sheep anti mouse polyclonal antibody control line are arranged on the nitrocellulose filter, in base plate one end termination is water accepting layer, other end termination is the glass fiber sample layer, the nitrocellulose filter two ends overlap mutually and are connected with anti-FSH, anti-LH monoclonal antibody gold mark pad with water accepting layer respectively, are pressed with the glass fiber sample layer on gold mark pad.

During detection, the sampling end of FSH, LH joint-detection test paper is put into detection sample (liquid level must not surpass the MAX line) because the capillarity test sample will move to the water accepting layer end along test strips, corresponding FSH, LH antigen can move and the antibodies that is coated on the nitrocellulose filter with continuation after anti-FSH monoclonal antibody gold mark probe, the anti-LH monoclonal antibody gold mark probe generation specific bond in the sample, thereby by anti-FSH, the identification of anti-LH monoclonal antibody, the specific bond of double antibodies sandwich takes place promptly.In the testing process, the gold mark probe that promptly is stranded in this zone when up liquid level will show corresponding redness, and it is positive two or above red line to occur, shows that FSH and LH level are higher than normally, might be disease of ovary; A red line occurs and be expressed as feminine gender.

When moving to anti-mouse polyclonal antibody control line, no matter FSH, LH content have or not in the sample, and gold mark probe all can combine delay with the sheep anti-mouse igg of bag quilt on the nitrocellulose filter, and it is red that control line is shown.So control line does not have, and colour band produces then representative operation wrong (during detection, the sample liquid level surpasses the MAX line) or reagent is expired.

Because adopt technique scheme, FSH provided by the present invention, LH joint-detection test paper have such beneficial effect: i.e. high specificity, highly sensitive, easily store, need not the technical skill personnel operation, do not need instrument and equipment, and readability as a result.

Description of drawings

Fig. 1 is the main TV structure figure of FSH, LH joint-detection test paper.

Fig. 2 is the plan structure figure of FSH, LH joint-detection test paper.

Fig. 3 is the positive findings figure of FSH, LH joint-detection test paper.

Fig. 4 is the negative findings figure of FSH, LH joint-detection test paper.

Among the figure 1, water sucting plate, 2, nitrocellulose filter, 3, sheep anti mouse polyclonal antibody control line, 4, test wire, 5 anti-FSH monoclonal antibodies gold mark pads, anti-LH monoclonal antibody gold mark pad, 6, the urine liquid-adsorption layer, 7, base plate, 8, the MAX line.

Specific embodiment

1. synthetic immunogen

(1) with the FSH immunogene (FSH-BSA) of the synthetic two kinds of different binding sites of chemical synthesis process;

(2) with the LH immunogene (LH-BSA) of the synthetic two kinds of different binding sites of chemical synthesis process.

2. MONOCLONAL ANTIBODIES SPECIFIC FOR

The FSH-BSA of two kinds of different binding sites that (1) will prepare is immune BALB/c mouse respectively.Set up the knurl strain with the SP20 Fusion of Cells and screen, get oncocyte and be injected in the BALB/c mouse abdominal cavity, make it produce ascites.Extract the screening of mouse ascites purifying, obtain the two kinds of FSH monoclonal antibody I, the FSH monoclonal antibody II that combine with FSH molecule different loci.

The LH-BSA of two kinds of different binding sites that (2) will prepare is immune BALB/c mouse respectively.Set up the knurl strain with the SP20 Fusion of Cells and screen, get oncocyte and be injected in the BALB/c mouse abdominal cavity, make it produce ascites.Extract the screening of mouse ascites purifying, obtain the two kinds of LH monoclonal antibody I, the LH monoclonal antibody II that combine with LH molecule different loci.

The preparation of collaurum and with the combining of progesterone monoclonal antibody

(1) get distilled water and add an amount of gold chloride magnetic agitation and be warmed to 90～95 ℃, add an amount of citric acid three and receive and continue heated and stirred to seething with excitement 5 minutes, it is standby to keep in Dark Place after the cooling.

(2) the collaurum liquid of FSH, LH labeling of monoclonal antibody, on the adsorbing fiber material, dry back is standby respectively.

4. film-making machine system film: utilize computer control transmission speed, guarantee that the antibody amount of bag quilt on the per unit film equates.

5. test strips combination: press the known technology combination.

6. using method: collect urina sanguinis, test paper liquid-adsorption layer end is inserted in the urine, the sample to be tested interface surpasses " MAX " line on the diagnose test paper, observations in 5 minutes.The colour developing district is two red line, and detection line is darker or equal than control line color, and then testing result is positive; It is negative when response line is shallower than control line or a control line only occurs; Colour band does not appear in control line, shows the expired or misoperation of test paper, please gets test paper in addition and detects again.

Claims

1. FSH, LH joint-detection test paper, it is characterized in that it is by base plate (7), water sucting plate (1), nitrocellulose filter (2), anti-FSH monoclonal antibody gold mark pad (5), anti-LH monoclonal antibody gold mark pad (5), sample liquid-adsorption layer (6), MAX line (8) is formed, the base plate middle part is a nitrocellulose filter, a FSH monoclonal antibody test wire (4) is arranged on the nitrocellulose filter,, a LH monoclonal antibody test wire (4) and a sheep anti mouse polyclonal antibody control line (3), in base plate one end termination is water sucting plate, other end termination is the sample liquid-adsorption layer, the nitrocellulose filter two ends respectively with water sucting plate and anti-FSH, anti-LH monoclonal antibody gold mark pad overlaps mutually and connects, and is pressed with the sample liquid-adsorption layer on gold mark pad.

2. a kind of test paper according to claim 1 is characterized in that being coated with on the described nitrocellulose filter three lines, two test wires, a control line.

3. a kind of test paper according to claim 1, it is characterized in that LH, FSH filter out the different monoclonal antibodies of two strains respectively by immunity, wherein a strain is used to wrap quilt, and another strain is used for colloid gold label, common and immune original affinity interaction separately, the test paper that utilizes double antibodies sandwich method principle to make.

4. test paper according to claim 1 is characterized in that the test wire color of utilizing the double antibodies sandwich method to make test paper is redness, and promptly colour developing district is two lines or positive when above; Negative when having only a control line for redness.