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Immune-chromatographic test strip for fluorescence quantitative detection of FSH (follicle stimulating hormone) and preparation method of immune-chromatographic test strip

A fluorescent quantitative detection and immunochromatographic test paper technology, applied in the field of immunodiagnosis, can solve the problems of many influencing factors, high price, long time consumption, etc., and achieve the effect of ensuring accuracy, simple operation and high sensitivity

Inactive Publication Date: 2017-09-15
SHENZHEN YHLO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ELISA method has many detection steps, takes a long time, and there are many influencing factors in the operation process, which can easily cause false positive and false negative results
Therefore, it is gradually replaced by chemiluminescence method. However, this method is a fully closed system, which is expensive and requires special training for instrument users. The cost of maintenance and testing is high, and it is not suitable for single-person and small-batch testing. It is not conducive to FSH at present. Testing is widely carried out in China. The colloidal gold method is simple to operate and suitable for single-person testing, but the test results can only be observed with the naked eye, giving qualitative or semi-quantitative results.
[0006] In view of the fact that there is no fast and accurate method for quantitatively detecting FSH at present, the purpose of the present invention is to provide an immunochromatographic test strip that can be used for rapidly quantitatively detecting FSH, for evaluating and monitoring the level of follicle-stimulating hormone in the body, so The above reagents have the advantages of simple operation, convenience, economy, accurate quantification, etc., and are more suitable for extensive use in various medical institutions.

Method used

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  • Immune-chromatographic test strip for fluorescence quantitative detection of FSH (follicle stimulating hormone) and preparation method of immune-chromatographic test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Preparation of FSH fluorescent immunochromatographic test strips:

[0035] (1) Preparation of fluorescent microsphere-labeled protein

[0036]Take 0.1 mL of 10% fluorescent microspheres, centrifuge at 15,000 rpm for 15 minutes, adjust the concentration of the precipitate to 1% with 50 mM pH 6.5 MES buffer, and ultrasonically disperse; add carbodiimide (EDC) at a final concentration of 2 mg / mL ), mix well, then add N-hydroxysuccinimide (NHS) with a final concentration of 5 mg / mL, mix well; incubate at room temperature for 20 minutes and then centrifuge at 15,000 rpm for 15 minutes, and the precipitate is washed with 50 mM pH6.5 MES buffer dissolve. Ultrasonic disperse the reconstituted fluorescent microspheres, and add 0.2 mg FSH monoclonal antibody and 0.1 mg avidin into two tubes respectively, mix well, rotate and mix at room temperature for 2 hours, centrifuge at 15000 rpm for 15 minutes, and deposit with 30 Reconstitute mM ethanolamine and 0.5% casein in Tris-HCl (...

Embodiment 2

[0045] Quantitative detection of follicle-stimulating hormone (FSH) concentration in blood samples by fluorescence immunochromatography

[0046] (1) Standard curve drawing

[0047] Prepare FSH antigen negative plasma to 150 mIU / mL, 75 mIU / mL, 30 mIU / mL, 15 mIU / mL, 5 mIU / mL, 1 mIU / mL, 0 mIU / mL, use the same batch of reagents, each Each concentration point was tested 6 times. Take the fluorescence intensity ratio of the detection line (T band) and the quality control line (C band) as the ordinate, and the concentration of FSH reference substance as the abscissa, establish an equation and fit it into a standard curve, and write the calibration information into In the ID chip.

[0048] (2) Detection of samples:

[0049] Take out the test strip from the kit, tear open the aluminum foil bag, lay the test strip flat, and equilibrate for 5 minutes. Take 100 μL of sample and add it to the sample well, and react at room temperature for 15 minutes in the dark. Insert the ID chip into...

Embodiment 3

[0053] Please refer to figure 1 , an immunochromatographic test strip for fluorescent quantitative detection of AMH, the immunochromatographic test strip includes a PVC base plate 7, and the PCV base plate 7 is sequentially provided with a sample pad 1, a marker pad 2, and a coating film 3. Absorbent paper 4, the marking pad 2 is connected to the sample pad 1, the coating film 3 is connected to the marking pad 2, the absorbent paper 4 is connected to the coating film 3 are connected; the FSH monoclonal antibody labeled with fluorescent microspheres and avidin labeled with fluorescent microspheres are sprayed on the marker pad 2; a detection line 5 and a quality control line 6 are provided on the coating film 3, and the detection line 5 and the quality control line 6 are separated by 4-8 mm, and the detection line 5 is coated with another FSH monoclonal antibody that is in a different epitope from the FSH monoclonal antibody labeled with the fluorescent microspheres, and the qu...

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Abstract

An immunochromatographic test strip for fluorescent quantitative detection of FSH and a preparation method thereof. The invention belongs to the technical field of immunodiagnosis. The purpose of the invention is to address the shortcomings in the prior art. The invention adopts the following technical solutions: The immunochromatographic test strip for quantitative detection of FSH is composed of a sample pad, a marking pad, a coating film, and an absorbent paper lapped on a PVC bottom plate in sequence. Compared with the prior art, the present invention has the following advantages: the detection line and the quality control line of the present invention adopt independent reaction systems, which do not interfere with each other and are calibrated by T / C value, which ensures the accuracy of test results Spend. The invention adopts fluorescence immunochromatography, and the detection method has high sensitivity, simple operation and low cost. One-step direct sample addition, without sample diluent, can detect follicle-stimulating hormone in blood samples with a concentration as low as 1.0 mIU / mL. The detector used does not require professional operators, and the test results can be obtained in 15 minutes.

Description

technical field [0001] The invention belongs to the technical field of immunodiagnosis, and in particular relates to an immunochromatographic test strip for fluorescent quantitative detection of follicle-stimulating hormone (FSH) and a preparation method thereof. Background technique [0002] Follicle stimulating hormone (FSH) is a glycoprotein hormone with two subunits. The alpha subunit is similar to that of luteinizing hormone (LH), human chorionic gonadotropin (hCG), and thyroid-stimulating hormone (TSH). The β subunit differs from that of other glycoprotein hormones and displays its specific biochemical properties. [0003] In women, FSH plays a role in the hypothalamus-pituitary-ovary regulatory loop, controlling the menstrual cycle. FSH and LH are released intermittently from the gonadotropic cells of the pituitary gland, and blood levels are controlled by steroid hormones through a negative feedback mechanism in the hypothalamus. Together, FSH and LH stimulate the...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/577G01N33/543G01N33/533
CPCG01N33/558G01N33/533G01N33/54313G01N33/577
Inventor 胡鹍辉夏福臻钱纯亘张赛宋永波
Owner SHENZHEN YHLO BIOTECH
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