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Immunomagnetic bead purification kit and method for 12 kinds of swine common viruses and germs

A technology of immunomagnetic beads and kits, which is applied in biochemical equipment and methods, bacteria, recovery/purification, etc., can solve the problem that the separation and purification conditions of biotinylated streptomycin magnetic beads have not been studied in detail, and it is not easy to achieve Effects of environmental impact, short operating time, and simple operation

Inactive Publication Date: 2016-07-20
BEIJING YISEN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies on the immunopurification and separation of various viruses and bacteria using immunomagnetic beads, and there is no detailed study on the separation and purification conditions of biotinylated streptomycin magnetic beads

Method used

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  • Immunomagnetic bead purification kit and method for 12 kinds of swine common viruses and germs
  • Immunomagnetic bead purification kit and method for 12 kinds of swine common viruses and germs
  • Immunomagnetic bead purification kit and method for 12 kinds of swine common viruses and germs

Examples

Experimental program
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Effect test

Embodiment 112

[0051] Embodiment 112 kinds of pig common viruses and bacterial immune magnetic bead purification method establishment

[0052] 1. Materials: Streptomycin Magnetic Beads (Cat. No. SVP-05-100) were purchased from Becton, Dickinson and Company, USA; porcine circovirus type II monoclonal antibody (Cat. No. ab183908), porcine pseudorabies virus monoclonal antibody (Cat. No. ab3534) , Streptococcus suis type II monoclonal antibody (Cat. No. ab251629), porcine parvovirus monoclonal antibody (Cat. No. ab140431), transmissible gastroenteritis virus monoclonal antibody (Cat. No. ab10024), epidemic diarrhea virus monoclonal antibody (Cat. No. ab161706) , rotavirus monoclonal antibody (product number ab130835), swine influenza virus monoclonal antibody (product number ab128412), swine fever virus monoclonal antibody (product number ab161081) and porcine foot-and-mouth disease virus monoclonal antibody (product number ab82724) were purchased from Abcam (Shanghai) Trading Co., Ltd.; porc...

Embodiment 2

[0109] Example 2: Preparation of Immunomagnetic Bead Purification Kit and Sample Detection of 12 Common Pig Viruses and Bacteria

[0110] 1. Preparation of the kit:

[0111] Reagent A: 1 ml (1 mg / ml) of streptomycin magnetic beads. (Becton, Dickinson and Company, USA)

[0112] Reagent B: biotinylated monoclonal antibody mixture 1 μg / μl, 25 μl.

[0113] Reagent C: 100ml buffer solution of 10× immunomagnetic bead separation and purification system, the preparation method is as follows:

[0114] Potassium dihydrogen phosphate (KH 2 PO 4 ): 0.27g

[0115] Disodium hydrogen phosphate (Na 2 HPO 4 ): 1.42g

[0116] Sodium chloride (NaCl): 8g

[0117] Potassium chloride (KCl): 0.2g

[0118] Bovine serum albumin (BSA): 3g

[0119] Add about 80ml of deionized water and stir well to dissolve, then add concentrated hydrochloric acid to adjust the pH to 7.4, and finally set the volume to 100ml. Store at 4°C after high temperature and high pressure sterilization. (It needs to...

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Abstract

The invention provides an immunomagnetic bead purification kit and method for 12 kinds of swine common viruses and germs. The kit comprises a reagent A, a reagent B and a reagent C, wherein the reagent A is streptomycin immunomagnetic suspension; the reagent B is a 12-biotinylation monoclonal antibody mixed solution; and the reagent C is a 10* immunomagnetic bead separation and purification system buffer solution. The method comprises the following steps of (1) performing a pathogen suspension preparation process; (2) performing a biotin antibody preparation process; and (3) performing a pathogen immunological purification process. The immunomagnetic bead purification kit and method for 12 kinds of swine common viruses and germs provided by the invention have the advantages that 12 kinds of viruses and germs such as PPV (Porcine Parvovirus), SS-II (Streptococcus Suis Type 2), PRV (Porcine Pseudorabies Virus) and the like in a sample can be simultaneously, fast and effectively purified. The detection method has the advantages of high accuracy, high specificity, high sensitivity and high stability, and the subsequent fast diagnosis and effective detection work is facilitated.

Description

technical field [0001] The invention relates to the field of biotechnology detection of bacteria and viruses, in particular to a kit and method for simultaneously separating and purifying 12 kinds of pig common viruses and bacteria by using immunomagnetic beads. Background technique [0002] my country is a large agricultural and animal husbandry country. Pig farming is mainly large-scale and intensive farming. The high-density breeding method also makes it difficult to prevent infectious diseases in pigs, and the problem is becoming more and more significant. Moreover, multiple pathogens are infected at the same time, which makes diagnosis and prevention of diseases very difficult. Porcine parvovirus (PPV), Streptococcus suis type 2 (SS-II), porcine pseudorabies virus (Porcinepseudorabies virus, PRV), porcine circovirus type II (Porcine circovirus- 2, PCV-2), transmissible gastroenteritis virus (Transmissiblegastroenteritisvirus, TEGV), epidemic diarrhea virus (Porcineepid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/02C12N1/20C12Q1/70C12Q1/68C12R1/93
Inventor 王新杰高姗姗孙晓明胡祥钰
Owner BEIJING YISEN BIOTECH
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