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Dot immuno gold directed infiltration detection kit and application thereof

A speckle immunization and detection technology, applied in the fields of scientific research, disease diagnosis and food safety, can solve the problems affecting detection sensitivity, loss of samples to be tested and labeled probes, etc., to improve detection sensitivity, suitable for large-scale promotion, and simple to manufacture. Effect

Inactive Publication Date: 2011-05-11
XIAN WEITONG BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that most of the samples to be tested and the labeled probes are lost from non-related areas other than the spots, which affects the detection sensitivity, so that this technology has not been widely used in clinical testing.

Method used

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  • Dot immuno gold directed infiltration detection kit and application thereof
  • Dot immuno gold directed infiltration detection kit and application thereof
  • Dot immuno gold directed infiltration detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Preparation of Dot Immunogold Directed Diafiltration Kit

[0042] The surface layer of the dot immunogold directional filtration card is white 5×4×0.5cm 3 The PVC card has a circular hole with a diameter of 0.8cm in the center; the microporous membrane of the percolation card is an NC membrane with a diameter of 1.2cm (the micropore is 0.45). Immobilize 1-2μl capture probe (1mg / ml) in the center of the microporous membrane using micro-spotting or spraying technology, in the form of circular spots (or strips) with a diameter of 2mm. After drying at room temperature or by air, immerse in 1% cattle Serum was dried for 1 minute at room temperature or air-dried, then room temperature or air-dried for 20 minutes; the percolation limiting layer of the diafiltration card was 5×4×0.5cm 3 Double-sided adhesive film, with a round hole (or strip) with a diameter of 2mm in the center, corresponding to the coated probe area in the center of the microporous membrane; the ab...

Embodiment 2

[0045] Example 2 Fabrication of Double Dot Immunogold Directed Diafiltration Card

[0046] The dual-spot immunogold directional filtration card displays the positive control and detection spots in the same detection well at the same time. The surface layer of the dot immunogold directional filtration card is white 5×4×0.5cm 3 The PVC card has a circular or oval opening with a diameter of 1cm in the center; the microporous membrane of the percolation card is an NC membrane with a diameter of 1.5cm (the micropore is 0.45). Fix 1-2μl of anti-gold probe antibody (1mg / ml) and capture probe (1mg / ml) in the center of the microporous membrane using micro-spotting or spraying techniques, respectively, with a distance of 4mm between them and a circle with a diameter of 2mm Type spots (or strips), at room temperature or after air-drying, immerse in 1% bovine serum for 1 minute, room temperature or air-drying and then room temperature or air-drying for 20 minutes; the percolation limitin...

Embodiment 3

[0047] Example 3 Production of Dot Immunogold Directed Diafiltration Card (Double Antibody Method) for Detecting Antigens

[0048] The surface layer of the dot immunogold directional filtration card is white 5×4×0.5cm 3 The PVC card has a circular hole with a diameter of 0.8cm in the center; the microporous membrane of the percolation card is an NC membrane with a diameter of 1.2cm (the micropore is 0.45). Immobilize 1-2μl of capture antibody (1mg / ml) in the center of the microporous membrane by using micro-spotting or spraying technology, in the form of circular spots (or linear and circular) with a diameter of 2mm. After drying at room temperature or by air, immerse in 1 % bovine serum for 1 minute, room temperature or air drying and then room temperature or air drying for 20 minutes; all the other operations are the same as in Example 1.

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Abstract

The invention relates to a dot immuno gold directed infiltration detection kit and application thereof. The kit comprises a dot immuno gold infiltration card, nano colloidal gold or a detection probe marked by latex microballoons, a negative standard product, a positive standard product and flushing liquor. The kit is technically characterized in that the dot immuno gold infiltration card is a dot immuno gold directed infiltration card which can be used for ensuring that a sample to be detected and the detection probe marked by the latex microballoons infiltrate sequentially along an area covered by a coated probe. The invention has the advantages of simpleness of manufacture, less reagent consumption, wide detection range and good maneuverability, and compared with a traditional dot immuno gold directed infiltration method, the detection sensitivity is obviously improved.

Description

technical field [0001] The invention relates to a dot immunogold directed diafiltration detection kit, which can be used in the fields of scientific research, disease diagnosis and food safety. Background technique [0002] Gold standard immunoassay technology is a new type of immunolabeling technology after isotope, fluorescein and enzyme labeling technology. The method is simple, fast, does not require special equipment, and has excellent visibility in result determination. It is suitable for 'bedside' detection, disease census and epidemic monitoring, as well as customs, food safety and aquaculture and other fields. [0003] Commonly used gold-labeled immunoassays include Bilateral immune-chromatographic assay and Dot immune-gold filtration assay. Lateral immunogold chromatography is a method in which the sample to be tested and the colloidal gold-labeled probe move laterally on a microporous membrane (for example: nitrocellulose membrane) by siphon action, and capture ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/576G01N33/569G01N33/571G01N33/574
CPCB82Y15/00C12Q1/6804G01N33/538C12Q2563/137C12Q2563/149G01N33/5436Y02A50/30
Inventor 林远廖帮琳
Owner XIAN WEITONG BIOSCI
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