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Chromatographic strip used for diagnosis of prostate cancer, and preparation method and application thereof

A technology for prostate cancer and chromatography test strips, which is applied in the directions of analysis materials, measuring devices, instruments, etc., can solve the problems of complicated methods for diagnosing prostate cancer and low accuracy, and achieves improved screening and diagnosis, simple operation and simple structure. Effect

Inactive Publication Date: 2018-11-13
SHANGHAI EAST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0012] Aiming at the above-mentioned technical problems in the prior art, the present invention provides a chromatography test strip for diagnosing prostate cancer and its preparation method and application, the said chromatography test strip for diagnosing prostate cancer The preparation method and application thereof shall solve the technical problems that the method for diagnosing prostate cancer in the prior art is complicated and the accuracy rate is not high

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  • Chromatographic strip used for diagnosis of prostate cancer, and preparation method and application thereof
  • Chromatographic strip used for diagnosis of prostate cancer, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Coupling of magnetic nanomaterials with total-PSA antibody

[0040] 1. Activation: Use pH 5.0, 0.01 M anhydrous morpholineethanesulfonic acid-Tween solution (MEST, 0.05% Tween-20) as the activation buffer solution, take 2 mg magnetic nanospheres in a 2 mL centrifuge tube, Add 500 μL activation buffer for washing: Sonicate and vortex to resuspend the magnetic beads in an ultrasonic cleaner, then perform magnetic separation with a magnetic separation stand for 5 minutes, remove the supernatant; add 500 μL activation buffer and repeat the same operation to wash the microspheres After two times, add 485 μl of activation buffer to the microspheres, add carbodiimide and 2.5 mg of N-hydroxysuccinimide at a concentration of 0.25 g / mL, and resuspend in an ultrasonic cleaner and vortex After mixing the beads, place them on a vertical mixer for 30 minutes of reaction at room temperature, then use a magnetic separation rack for magnetic separation for 5 minutes, remove th...

Embodiment 2

[0044] Example 2 Construction of joint detection test strips

[0045] The present invention provides a chromatographic test strip for diagnosing serum markers of prostate cancer, comprising a bottom plate 7, on which a sample pad 1, a binding pad 2, a blood barrier layer, a chromatographic A pad 8 and a water-absorbing pad 6, the binding pad 2 is provided with a magnetic nano-immune probe coupled with total-PSA antibody; the chromatography pad 2 is sequentially provided with a first detection line 3 according to the direction of liquid flow , the second detection line 4 and the quality control line 5, the first detection line 3 is provided with an anti-freePSA monoclonal antibody, and the second detection line 4 is provided with a prostate-specific anti-alpha-1 anti-chymotrypsin ACT For antigen antibodies, the quality control line 5 is provided with IgG antibodies capable of binding to total PSA antibodies.

[0046] Further, a blood-separating layer is also included, and the ...

Embodiment 3

[0054] Example 3 Sample detection and result analysis

[0055] 1. Take 10 μL of magnetic microsphere dispersion coupled with total PSA antibody, mix it with 20 μL of chromatography solution (PBS with 1% calf serum) and 100 μL of the solution to be tested, take 100 μL with a pipette A liquid gun is added to the sample pad for testing.

[0056] 2. After adding the sample for 10 minutes, according to figure 2 , first check whether there is a band on the quality control line to determine the validity of the test strip, and then use the instrument to test the magnetic signal value of the first detection line and the second detection line to achieve quantitative detection; put the test strip into the MAR instrument Carry out magnetic detection to obtain the magnetic signal values ​​of T1, T2, and C lines, and judge the content of free-PSA and PSA-ACT of the sample to be tested according to the self-made standard curve. The result analysis is based on the sum of the concentrations...

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Abstract

The invention discloses a chromatographic strip used for diagnosis of prostate cancer. The chromatographic strip comprises a bottom plate; a sample pad, a conjugate pad, a blood insulation layer, a chromatographic pad and a water absorption pad are sequentially arranged on the bottom plate in an overlapping manner; a magnetic nanometer immunoprobe coupled with an anti-total prostate cancer specific antigen-antibody is arranged on the conjugate pad; a first detecting line, a second detecting line and a quality control line are sequentially arranged on the chromatographic pad; a monoclonal antibody with anti-free prostate cancer specific antigen is arranged in the first detecting line, PSA(prostate-specific antigen)-antibody combined with anti-Alpha-1 antichymotrypsin (ACT) is arranged on the second detecting line, and IgG antibody capable of being combined with total PSA antibody is arranged on the quality control line. The invention further provides a preparation method of the strip. According to the invention, the occurrence of the problem of incapability of determining the ratio of two antigens, caused by partial overlapping of detecting sites between free PSA and total PSA whichare preset on the detecting line originally, is greatly reduced, and the accuracy of the detecting results are improved.

Description

technical field [0001] The invention belongs to the field of biological detection and relates to a test strip, in particular to a chromatographic test strip for diagnosing serological markers of prostate cancer and its preparation method and application. Background technique [0002] Lateral Flow ImmunoAssay (LFIA) is a method that has attracted widespread attention in recent years. This technology has developed rapidly in recent years. It is based on the immune sandwich reaction, which can realize rapid, specific and highly sensitive detection of targets on nitrocellulose test strips. At present, methods such as colloidal gold immunochromatography, immunoenzyme staining chromatography, and magnetic immunochromatography have been developed. In the future, immunochromatography will move towards quantitative, high-sensitivity, multiple detection, and chromatography system integration. develop. This technology has the advantages of being fast, simple, and does not require co...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/558
CPCG01N33/558G01N33/57434
Inventor 王祎龙杨昊成
Owner SHANGHAI EAST HOSPITAL
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