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Hepatocyte growth factor detection and signal sensitization probe, primer pair and application thereof

A hepatocyte growth factor and growth factor technology, which is applied in biological testing, biochemical equipment and methods, and microbial measurement/inspection, etc., can solve the problems of high detection conditions and achieve good specificity, simple operation, and high sensitivity Effect

Pending Publication Date: 2022-04-08
THE FIRST AFFILIATED HOSPITAL OF ANHUI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The patent application with the publication number CN112326956A discloses a tumor-specific growth factor latex-enhanced immune turbidimetric detection kit, which has high sensitivity and specificity, but it needs to use a fully automatic biochemical analyzer in the detection process, which is very difficult for detection. higher requirements

Method used

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  • Hepatocyte growth factor detection and signal sensitization probe, primer pair and application thereof
  • Hepatocyte growth factor detection and signal sensitization probe, primer pair and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] The method for rapid detection of hepatocyte growth factor by lateral flow immunochromatography specifically comprises the following steps:

[0073] (1) FITC antibody, HGF antibody 1 and colloidal gold coupling

[0074] Adjust pH: Take 1000 μL of red colloidal gold with a particle size of 40 nm in a 1.5 mL centrifuge tube, add 18 μL of 0.1mol / L K 2 CO 3 Mix the aqueous solution;

[0075] Coupling: Add 1.5 μL of 1 mg / mL FITC antibody and 1 mg / mL HGF antibody 1 respectively, vortex and mix well, and incubate at room temperature for 1 h.

[0076] Blocking: Add 100 μL of 1×PBS solution with a mass volume ratio of 10% BSA, vortex to mix, and incubate at room temperature for 1 h.

[0077] Resuspension: Centrifuge at 8500g, 4°C, 15min, remove the supernatant, then redissolve the pellet in 100μL of 1% BSA in 1×PBS solution, store at 4°C, and obtain FITC antibody and HGF antibody 1-labeled colloidal gold solution.

[0078] (2) The FITC-modified detection probe binds to the ...

Embodiment 2

[0100] The method for rapid detection of hepatocyte growth factor by lateral flow immunochromatography specifically comprises the following steps:

[0101] (1) FITC antibody, HGF antibody 1 and colloidal gold coupling

[0102] Adjust pH: Take 1000 μL of red colloidal gold with a particle size of 18nm in a 1.5mL centrifuge tube, add 18μL of 0.1mol / L K 2 CO 3 Mix the aqueous solution;

[0103] Coupling: Add 1.5 μL of 1 mg / mL FITC antibody and 1 mg / mL HGF antibody 1 respectively, vortex and mix well, and incubate at room temperature for 1 h.

[0104] Blocking: Add 100 μL of 1×PBS solution with a mass volume ratio of 10% BSA, vortex to mix, and incubate at room temperature for 1 h.

[0105] Resuspension: Centrifuge at 8500g, 4°C, 15min, remove the supernatant, then redissolve the pellet in 100μL of 1% BSA in 1×PBS solution, store at 4°C, and obtain FITC antibody and HGF antibody 1-labeled colloidal gold solution.

[0106] (2) The FITC-modified probe binds to the FITC antibody...

Embodiment 3

[0127] The method for rapid detection of hepatocyte growth factor by lateral flow immunochromatography specifically comprises the following steps:

[0128] (1) FITC antibody, HGF antibody 1 and colloidal gold coupling

[0129] Adjust pH: Take 1000μL of red colloidal gold with a particle size of 20nm in a 1.5mL centrifuge tube, add 18μL of 0.1mol / L K 2 CO 3 Mix the aqueous solution;

[0130] Coupling: Add 2 μL of 1 mg / mL FITC antibody and 1 mg / mL HGF antibody 1 respectively, vortex and mix well, and incubate at room temperature for 1 h.

[0131] Blocking: add 100 μL of 1×PBS solution with a mass volume ratio of 15% BSA, vortex to mix, and incubate at room temperature for 1 h.

[0132] Resuspension: centrifuge at 9500g, 4°C, 15min, remove the supernatant, then redissolve the pellet in 100 μL 0.5% BSA in 1×PBS solution, and store at 4°C.

[0133] (2) The FITC-modified detection probe binds to the FITC antibody on the colloidal gold that labels the two antibodies

[0134] Take ...

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Abstract

The invention discloses a hepatocyte growth factor detection and signal sensitization probe, and relates to the technical field of hepatocyte growth factor detection, the 5'end of the probe is modified with fluorescein (FITC), and the sequence of the probe is shown as SEQ ID NO.1. The invention also provides a primer pair for amplifying the probe by adopting an enzymatic recombination isothermal amplification method and application of the primer pair. The method has the beneficial effects that a target probe is indirectly captured by utilizing a sandwich method, and an enzymatic recombinant isothermal amplification (ERA) technology and a lateral immunochromatography technology are combined to amplify a detection signal, so that the detection sensitivity is improved.

Description

technical field [0001] The invention relates to the technical field of hepatocyte growth factor detection, in particular to a hepatocyte growth factor detection and signal sensitization probe, a primer pair and applications thereof. Background technique [0002] Heart failure is a clinical syndrome in which symptoms and / or signs are caused by structural and / or functional abnormalities of the heart. It has become a global epidemic in the 21st century with increasing impact on the health care system. According to statistics, the global age-standardized prevalence rate of heart failure in 2017 was 0.83%, which was 7.2% lower than that in 1990, but the absolute number of heart failure patients nearly doubled, and 90% of the increase came from China. According to the China Hypertension Survey, the weighted prevalence of heart failure among residents aged ≥35 years in my country was 1.3% in 2015, an increase of 44.0% compared with 2000. These data all show that the prevalence of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6804C12N15/11G01N33/68G01N33/543G01N33/558G01N33/58
Inventor 胡泽平陈琦冯静陈伟朱学涛
Owner THE FIRST AFFILIATED HOSPITAL OF ANHUI MEDICAL UNIV
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