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Liquid-phase protein chip kit for simultaneously detecting four types of foreign and new animal diseases and preparation method thereof

A protein chip and kit technology, which is applied in the field of liquid-phase protein chips for simultaneous detection of multiple animal diseases, can solve problems such as the inability to realize real-time monitoring of multiple pathogenic antibodies, and achieve good specificity and sensitivity, short time and operation. simple effect

Inactive Publication Date: 2018-05-04
贾赟
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have a common disadvantage: one test can only detect the antibody level of one pathogen, and it is impossible to realize real-time monitoring of antibodies to multiple pathogens

Method used

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  • Liquid-phase protein chip kit for simultaneously detecting four types of foreign and new animal diseases and preparation method thereof
  • Liquid-phase protein chip kit for simultaneously detecting four types of foreign and new animal diseases and preparation method thereof
  • Liquid-phase protein chip kit for simultaneously detecting four types of foreign and new animal diseases and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Preparation of a liquid-phase protein chip kit for simultaneous detection of four exotic and emerging animal diseases

[0039] 1. Preparation of capture microspheres

[0040] (1) Microsphere activation

[0041] Select four kinds of carboxyl fluorescent magnetic microspheres with the same diameter and material but with different codes (the commodity information is respectively Bio-Plex: MC10035-01; MC10047-01; MC10055-01 and MC10065-01, wherein 35, 47, 55 and 65 represent magnetic bead number), as the first microsphere, the second microsphere, the third microsphere and the fourth microsphere described in the present invention.

[0042] Select the selected microspheres (about 6.1×10 6 Each sample / mL) were subjected to the following operations: Vortex for 30 seconds, ultrasonically oscillate for 30 seconds; take 100 μL microbeads into the reaction tube, centrifuge at 14000×g for 4 minutes, carefully remove the supernatant; add 100 μL microbead washing solution, Vorte...

Embodiment 2

[0059] Using the liquid-phase protein chip kit of Example 1, 60 samples were simultaneously detected for four kinds of foreign and emerging animal diseases (source of samples: 20 copies of Dalian Animal Disease Prevention and Control Center, 15 copies of Inspection and Quarantine Technology Center of Yunnan Entry-Exit Inspection and Quarantine Bureau) 15 copies, 15 copies from the Animal and Plant Inspection and Quarantine Technology Center of the Shenzhen Entry-Exit Inspection and Quarantine Bureau, and 10 copies from the Chinese Academy of Inspection and Quarantine), including the following steps:

[0060] (1) Take out the capture microspheres provided in the kit, suspend them and add 50 μL to the 96-well experimental plate; the number of each type of microspheres in each well is about 5000;

[0061] (2) Add 50 μL of 1:1000 diluted quality control substance and 60 samples of serum to be tested in the corresponding wells (at least two replicates for each sample), and shake at ...

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Abstract

The invention discloses a liquid-phase protein chip kit for simultaneously detecting four types of foreign and new animal diseases. Four types of activated fluorescent magnetic microspheres are respectively coupled with the expressed, purified and verified peste des petits ruminants virus protein (PPRV-N), vesicular stomatitis virus protein (VSV-N), schmallenberg virus protein (SBV-N) and bluetongue virus protein (BTV-VP7). The liquid-phase protein chip kit has the characteristics that the four types of foreign and new animal diseases, such as peste des petits ruminants, vesicular stomatitis,schmallenberg and bluetongue, can be simultaneously detected; the parallel detection effect is realized, and the flux is high; about 400 items of 100 samples can be completed each time; the operationis simple, the time is short, and the detection can be completed within 1h or above; the usage amount of samples is small, and multiple items can be detected by only using 0.1 to 1mu L of serum; the specificity and sensitivity are good.

Description

technical field [0001] The invention relates to the technical field of liquid-phase protein chips, in particular to the liquid-phase protein chip technology for simultaneous detection of various animal diseases. Background technique [0002] Peste des petits ruminants (PPR), commonly known as sheep plague, also known as pseudorinderpest in small ruminants and pneumonia, is a peste des petits ruminants virus (Peste des petits ruminants) belonging to the genus Morbolivirus of the family Paramyxoviridae. des petits ruminants Virus (PPRV) is an acute, febrile and highly contagious severe infectious disease, mainly infecting goats and sheep, characterized by fever, stomatitis, diarrhea, and pneumonia. Cattle are invisible and will not infect other animals after infection. After the onset of sheep, the prognosis is generally poor, and most of them die. The symptoms and pathological changes of the disease and rinderpest are highly similar. [0003] Vesicular stomatitis (VS), als...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543G01N33/532G01N33/533
CPCG01N33/6893G01N33/532G01N33/533G01N33/54326
Inventor 贾赟张雪刘钊张永宁
Owner 贾赟
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