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Colloidal gold lateral flow test strip for breast cancer detection and colloidal gold lateral flow test strip for simultaneously detecting breast cancer and cervical cancer

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

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

AI Technical Summary

Problems solved by technology

Although these methods have a high detection rate, they are not only invasive and uncomfortable for the examiner, but the specificity and sensitivity of the detection are insufficient, and the diagnostic efficiency of the detection is low.

Method used

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  • Colloidal gold lateral flow test strip for breast cancer detection and colloidal gold lateral flow test strip for simultaneously detecting breast cancer and cervical cancer
  • Colloidal gold lateral flow test strip for breast cancer detection and colloidal gold lateral flow test strip for simultaneously detecting breast cancer and cervical cancer
  • Colloidal gold lateral flow test strip for breast cancer detection and colloidal gold lateral flow test strip for simultaneously detecting breast cancer and cervical cancer

Examples

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preparation example Construction

[0044] The preparation method of T1 line solution or T2 line solution of the present invention comprises the following steps: mixing biotin-labeled miR-16 partial complementary sequence or miR-191 partial complementary sequence with streptavidin solution and culturing to obtain T1 line solution or T2 line solution.

[0045] 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.

[0046] In the present invention, the T3 thread is sprayed with a T3 thread solut...

Embodiment 1

[0090] Example 1 A colloidal gold lateral flow chromatography test strip for breast cancer detection and its use

[0091] 1. Antibody-labeled colloidal gold: Take 1ml of five-fold concentrated nano-gold (15-20nm) solution, add 10μL of 1mM antibody (CA125 antibody, CA153 antibody, secondary antibody), shake with a shaker for 60min at room temperature, and then add 100μL of 10 %BSA, shake culture for 30min, centrifuge with a centrifuge (12,000rpm, 10min), remove the supernatant, wash with PBS buffer (pH7.2~7.4) for 3 times, and finally dissolve the precipitate in 1mL nanoparticle storage solution (20mmol / LNa 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.

[0092]2. DNA sequence labeling colloidal gold: take 1ml of ten-fold concentrated nano-gold (15-20nm) solution, add 10 μL of 1mM dATP, shake with an oscillator for 20 minutes at room temperature, then add 15 μL of 1% SDS, shake and incubate for 10 m...

Embodiment 2

[0098] Example 2 A method for preparing and using a colloidal gold lateral flow test strip for simultaneous detection of breast cancer and cervical cancer

[0099] 1. Antibody-labeled colloidal gold: Take 1ml of five-fold concentrated nano-gold (15-20nm) solution, add 10μL of 1mM antibody (CA125 antibody, CA153 antibody, E6 antibody, E7 antibody, secondary antibody), at room temperature, use a shaker Shake 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, wash 3 times with PBS buffer (pH 7.2-7.4), and finally dissolve the precipitate. In 1mL nanoparticle storage solution (20mmol / LNa 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.

[0100] 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 oscil...

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Abstract

The invention provides a colloidal gold lateral flow test strip for breast cancer detection, a preparation method of the colloidal gold lateral flow test strip and a colloidal gold lateral flow test strip for simultaneously detecting breast cancer and cervical cancer and belongs to the technical field of immuno-chemical detection. The colloidal gold lateral test strip comprises a PVC bottom plate,a nitrocellulose film, a sample pad, a conjugate pad and an absorption pad, and can be used for detecting miRNA markers (miR-16 and miR-191) and protein markers (CA153 and CA125) of breast cancer. The invention further provides a colloidal gold lateral test strip for simultaneously detecting breast cancer and cervical cancer, which can overcome the interference (hybridization among components) ofa background signal to realize specific detection on two different types of tumor marker groups, including miRNA groups and protein, of the cervical cancer and breast cancer, and has the detection specificity of 100 percent.

Description

technical field [0001] The invention relates to the technical field of immunochemical detection, in particular to a colloidal gold lateral chromatography test strip for detecting breast cancer and a colloidal gold lateral chromatography test strip for simultaneously detecting breast cancer and cervical cancer. Background technique [0002] Breast cancer is a malignant tumor that occurs in the glandular epithelial tissue of the breast. At present, the examination of breast cancer mainly relies on several methods such as physical examination, imaging examination (B-ultrasound, mammography X-ray, MRI), and histological cytology examination. Although these methods have a high detection rate, they are not only invasive and uncomfortable for examiners, but also have insufficient specificity and sensitivity of detection, and low diagnostic efficiency of detection. Contents of the invention [0003] The purpose of the present invention is to provide a colloidal gold lateral chrom...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6834G01N33/58G01N33/577G01N33/574G01N33/558
CPCC12Q1/6886C12Q1/6834G01N33/587G01N33/577G01N33/57415G01N33/57411G01N33/57484G01N33/558C12Q2600/158C12Q2600/178C12Q2525/207C12Q2565/625
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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