Fluorescence quantitative immunochromatography test paper strip for fast detecting zika virus NS1 protein and manufacturing method thereof

An immunochromatographic test strip and Zika virus technology, applied in the biological field, can solve the problems of high detection cost, difficulty in rapid screening of large-scale populations, investigation and research of past infection history, time-consuming detection of personnel, etc., to achieve accurate on-site Detection effect

Inactive Publication Date: 2017-08-29
SHENZHEN ZIJIAN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The detection cost of this method is high, it takes a long time, and it has certain qualification requirements for the detection personnel. It is difficult to apply to the rapid screening of large-sca

Method used

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  • Fluorescence quantitative immunochromatography test paper strip for fast detecting zika virus NS1 protein and manufacturing method thereof
  • Fluorescence quantitative immunochromatography test paper strip for fast detecting zika virus NS1 protein and manufacturing method thereof
  • Fluorescence quantitative immunochromatography test paper strip for fast detecting zika virus NS1 protein and manufacturing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1, as figure 1 As shown, a fluorescent quantitative immunochromatographic test strip for rapid detection of Zika virus NS1 protein includes a bottom plate 5 on which a sample pad 1, a binding pad 2, a coating film 3 and an absorbent paper 4 are sequentially arranged, And the sample pad 1, the binding pad 2, the coating film 3 and the absorbent paper 4 are overlapped sequentially; the binding pad 2 contains the Zika NS1 protein monoclonal antibody labeled with fluorescent microspheres and the goat anti-chicken antibody labeled with fluorescent microspheres. Antibody;

[0032]The coating membrane 3 includes a detection area T and a quality control area C, and the detection area T is coated with a Zika NS1 protein having a different epitope from the Zika NS1 protein monoclonal antibody labeled with fluorescent microspheres on the binding pad 2 Monoclonal antibody, the quality control region C is coated with chicken IgY antibody.

[0033] In the fluorescent qua...

Embodiment 2

[0047] Embodiment 2, a kind of preparation method of fluorescence quantitative detection Zika NS1 protein immunochromatography test strip, comprises the following steps:

[0048] 1. Cleaning and activation of fluorescent microspheres:

[0049] Fluorescent microspheres with a particle size of 500nm were selected, and the wavelengths of excitation light and emission light of the fluorescent microspheres were 470nm and 525nm, respectively. Centrifuge the fluorescent microsphere suspension at 12000×g for 10 min, remove the supernatant, and resuspend the precipitate with 20 mM morpholine ethanesulfonic acid solution at pH 5.0, and then treat it with 100 W ultrasonic wave for 50 seconds, so that the concentration of the fluorescent microsphere suspension is 10mg / ml, add 50μl carbodiimide and N-hydroxysulfosuccinimide solution in turn, place in a rotary mixer, mix at room temperature for 15min, centrifuge at 12000×g for 20min, discard After clearing, resuspend with 20mM morpholineet...

Embodiment 3

[0061] Embodiment 3, a kind of preparation method of fluorescence quantitative detection Zika NS1 protein immunochromatography test strip, comprises the following steps:

[0062] 1. Cleaning and activation of fluorescent microspheres:

[0063] Fluorescent microspheres with a particle size of 0.2 μm are selected, and the wavelengths of excitation light and emission light of the fluorescent microspheres are 400 nm and 750 nm, respectively. Centrifuge the fluorescent microsphere suspension at 12000×g for 20min, remove the supernatant, and resuspend the precipitate with 20mM morpholineethanesulfonic acid solution, pH 5.0-6.5, and treat with ultrasonic waves at 300W for 20s, so that the final concentration of fluorescent microspheres is 10mg / ml , add 100μl of 50mg / ml carbodiimide and N-hydroxysulfosuccinimide solution in turn, place in a rotary mixer, mix at room temperature for 30min, then centrifuge at 15000×g for 10min, discard the supernatant and wash with 50mM pH6 .5 morpholi...

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Abstract

The invention discloses a fluorescence quantitative immunochromatography test paper strip for fast detecting zika virus NS1 protein and a manufacturing method thereof. The test paper strip comprises a bottom plate, wherein a sample pad, a combination pad, a covering film and water suction paper are sequentially arranged on the bottom plate and are in sequential lap joint; a fluorescent microsphere marked zika NS1 protein monoclonal antibody and a fluorescent microsphere marked goat anti-chicken antibody are arranged on the combination pad; a zika NS1 protein monoclonal antibody with different epitope from the fluorescent microsphere marked zika NS1 protein monoclonal antibody on the combination pad covers a detection region of the covering film; a chicken IgY antibody covers a quality control region. The manufacturing method comprises the following steps of fluorescent microsphere cleaning and activation, fluorescent microsphere marked antibody preparation, fluorescent marked antibody combination pad preparation, covering film preparation and test paper strip assembly. The manufactured test paper strip can be used for fast, specially and precisely detecting the zika virus NS1 protein; the fast quantitative detection in field can be realized.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a fluorescent quantitative immunochromatographic test strip for viruses and a manufacturing method thereof, in particular to a fluorescent quantitative immunochromatographic test strip for rapidly detecting Zika virus NS1 protein and a manufacturing method thereof. Background technique [0002] Zika virus (Zika Virus) belongs to Flaviviridae (Flaviviridae), Flaviviridae (Flavivirus genus), is an arbovirus mainly transmitted by Aedes mosquitoes. The virus was first discovered in Uganda in 1947, and sporadic cases or small-scale outbreaks have continued to be found for many years since then. However, since 2015, human infections caused by Zika virus have continued to occur. A total of 32 countries and regions in the Americas, the Western Pacific, Africa and Asia have reported local transmission of Zika virus. [0003] At present, Zika virus is mainly detected by fluorescent PCR method, ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/569G01N33/558G01N33/543G01N33/533
CPCG01N33/68G01N33/533G01N33/54313G01N33/558G01N33/56983G01N33/577
Inventor 郑志于洪波
Owner SHENZHEN ZIJIAN BIOTECH
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