Immune-chromatographic test strip for fluorescence quantitative detection of INHB (inhibin B) 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 the chemiluminescence method, but 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 currently not conducive to INHB Exte

Method used

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  • Immune-chromatographic test strip for fluorescence quantitative detection of INHB (inhibin B) and preparation method of immune-chromatographic test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Preparation of INHB fluorescent immunochromatographic test strips:

[0032] (1) Preparation of fluorescent microsphere-labeled protein

[0033] 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 phosphate buffer, and ultrasonically disperse; add carbodiimide at a final concentration of 2 mg / mL ( EDC), 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 centrifuge at 15,000 rpm for 15 minutes, and the precipitate is washed with 50 mM pH6.5 phosphoric acid Saline buffer dissolves. Ultrasonic disperse the reconstituted fluorescent microspheres, and add 0.2 mg INHB monoclonal antibody and 0.1 mg avidin into two tubes respectively, mix well, rotate and mix at room temperature for 2 hours, and centrifuge at 15,000 rpm for 15 minutes. 30 mM ethanolamine and 0.5% casein in Tris-HCl (20...

Embodiment 2

[0042] Quantitative detection of inhibin B (INHB) concentration in blood samples by fluorescence immunochromatography

[0043] (1) Standard curve drawing

[0044] Prepare INHB antigen negative plasma to 1600 pg / mL, 800 pg / mL, 400 pg / mL, 100 pg / mL, 50 pg / mL, 25 pg / mL, 0 pg / mL, using the same batch of reagents, 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 INHB 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.

[0045] (2) Detection of samples:

[0046] 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 the fluorescence de...

Embodiment 3

[0050] 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 INHB 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 to 8 mm, and the detection line 5 is coated with another INHB monoclonal antibody that is at a different epitope from the INHB monoclonal antibody labeled with the fluorescent microspheres, and ...

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Abstract

An immunochromatographic test strip for fluorescence quantitative detection of INHB and a preparation method thereof. The invention belongs to the technical field of immunodiagnosis. The purpose of the invention is to address the deficiencies in the prior art. The invention adopts the following technical solutions: a fluorescent The immunochromatographic test strip for quantitative detection of INHB is composed of a sample pad, a marker pad, a coating film, and an absorbent paper sequentially lapped on a PVC bottom plate. The marker pad is sprayed with avidin; the coating of the quality control line has specific identification Rabbit anti-avidin antibody for avidin. The invention has the following advantages: the detection line and the quality control line adopt an independent reaction system without mutual interference and influence, and adopt the T/C value method for calibration, which ensures the accuracy of the test results. Fluorescence immunochromatography is adopted, and the detection method has high sensitivity, simple operation and low cost. It can detect inhibin B in blood samples with a concentration as low as 10 pg/mL. The detector used does not require professional operators, and the detection result 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 inhibin B (INHB) and a preparation method thereof. Background technique [0002] Inhibin B (inhibin B, INH-B) is a glycoprotein hormone derived from the testis. The serum level of inhibin B in adult males is significantly negatively correlated with FSH, which acts as a negative feedback on FSH. Shortly after birth in males, serum inhibin B levels gradually increased, and a negative correlation between inhibin B and FSH was maintained. At the age of 20 to 30, the level of inhibin B reaches its peak, and then the level of inhibin B gradually decreases with age. [0003] The level of serum inhibin B in men with low spermatogenic function and spermatogenic block was significantly lower than that in men with normal spermatogenic function, but the serum inhibin B level in men with Sertoli...

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 胡鹍辉钱纯亘夏福臻张赛宋永波
Owner SHENZHEN YHLO BIOTECH
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