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Colloidal gold lateral chromatography test strip for detecting cervical cancer and preparation method thereof

A technology of lateral flow chromatography and colloidal gold, which is applied in the field of immunochemical detection, can solve the problems of single detection object and insufficient diagnostic efficiency, and achieve high sensitivity, good diagnostic efficiency and high specificity

Active Publication Date: 2020-07-10
ANHUI SCI & TECH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above-mentioned prior art only detects protein tumor markers, the detection object is single, and the diagnostic efficiency of detection is insufficient

Method used

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  • Colloidal gold lateral chromatography test strip for detecting cervical cancer and preparation method thereof
  • Colloidal gold lateral chromatography test strip for detecting cervical cancer and preparation method thereof

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preparation example Construction

[0038] The preparation method of T1 line solution or T2 line solution of the present invention comprises the following steps: mixing biotin-labeled miR-21 partial complementary sequence or miR-196a partial complementary sequence with streptavidin solution and culturing to obtain T1 line solution or T2 line solution.

[0039] During the specific implementation of the present invention, 50 nM biotin-labeled T1 / T2 and 200 μL 2.5 mg / ml streptavidin (Shanghai Sangong) were mixed and cultured in 0.01 MPBS for 1 h, and then the mixed solution was transferred to a dialysis tube ( MWCO 30000), centrifuge in a refrigerated centrifuge (6000rpm, 20min, 4°C) to remove unbound aptamer probes. Add PBS and repeat the above steps twice, and finally collect the centrifuged solution and dilute to 600 μL. Finally, the solution of biotin-labeled DNA and streptavidin-conjugated mixture was sprayed on the test strip.

[0040] In the present invention, the T3 thread is sprayed with a T3 thread solu...

Embodiment 1

[0070] 1. Antibody-labeled colloidal gold: take 1ml of five-fold concentrated nano-gold (15-20nm) solution, add 10μL of 1mM antibody (mouse anti-HPV E6 protein monoclonal antibody, mouse anti-HPV E7 protein monoclonal antibody, secondary antibody), in At room temperature, shake with a shaker for 60 minutes, then add 100 μL of 10% BSA, shake and incubate for 30 minutes, centrifuge with a centrifuge (12,000 rpm, 10 minutes), remove the supernatant, and wash with PBS buffer (pH 7.2-7.4) for 3 Finally, dissolve the precipitate in 1 mL of nanoparticle storage solution (20 mmol / L Na 3 PO 4 12H 2 O, 5% BSA, 0.25% Tween 20, 10% sucrose), the conjugate solution was stored in a refrigerator at 4°C until use.

[0071] 2. Colloidal gold labeled with DNA detection probe: take 1ml of ten-fold concentrated nano-gold (15-20nm) solution, add 10μL of 1mM dATP, shake with an oscillator for 20min at room temperature, then add 15μL of 1% SDS, and incubate with shaking for 10min Finally, add 50μ...

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Abstract

The invention provides a colloidal gold lateral chromatography test strip for detecting cervical cancer and a preparation method thereof, and belongs to the technical field of immunochemical detection. The colloidal gold lateral chromatography test strip comprises a PVC base plate, a nitrocellulose membrane, a sample pad, a combination pad and an absorption pad. The colloidal gold lateral chromatography test paper provided by the invention can overcome the interference of background signals (hybridization reaction among components), and can specifically detect cervical cancer miRNA markers (miR-21, miR-196a) and protein markers (HPV E6, HPV E7) at the same time. Compared with a PCR technology and an enzyme-linked immunoassay technology which are generally adopted in the market at present,the colloidal gold lateral chromatography test strip for detecting cervical cancer has better detection efficiency.

Description

technical field [0001] The invention relates to the technical field of immunochemical detection, in particular to a colloidal gold lateral flow chromatography test strip for detecting cervical cancer and a preparation method thereof. Background technique [0002] Cervical cancer is the most common gynecological malignancy. At present, screening methods for cervical cancer mainly include VIA / VILI (Visual inspectT1on with aceT1c acid and iodine, visual inspection with acetic acid / iodine staining), cervical cytology, colposcopy, etc., but these methods cannot identify HPV infection The risk of genotype, simultaneous detection requires specific instruments and specialized technicians, which is not suitable for early and rapid screening of cervical cancer. The OncoE6 oncoprotein detection technology (AVC, Sunnyvale, CA, USA) developed by ArborVita in the United States is a capture experiment in the form of "test strip detection", which can detect HPV 16 and 18 E6 oncoproteins wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/558
CPCG01N33/57411G01N33/57484G01N33/558
Inventor 刘国东邱万伟钱立生张静李坤余庆才
Owner ANHUI SCI & TECH UNIV
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