Viral biological magnetic bead enrichment and concentration kit and its application

A biomagnetic and virus technology, applied in the directions of microorganisms, viruses/phages, biochemical equipment and methods, etc., can solve the problems of requiring centrifugation steps, inconvenient operation, difficult clinical application, etc., achieving simple operation, low cost, avoiding repeated centrifugal effect

Active Publication Date: 2021-07-20
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the existing magnetic bead enrichment detection method, its cost is high, the operation is inconvenient, and centrifugation steps are required, so it is difficult to be widely used in clinical practice

Method used

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  • Viral biological magnetic bead enrichment and concentration kit and its application
  • Viral biological magnetic bead enrichment and concentration kit and its application
  • Viral biological magnetic bead enrichment and concentration kit and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation of Virus Biomagnetic Beads Enrichment and Concentration Kit

[0027] The virus biological magnetic bead enrichment and concentration kit provided in this embodiment contains balance solution and Fe 3 o 4 nano magnetic beads.

[0028] The composition of the balance solution is as follows: glucose 200g / L, mannose 100g / L, galactose 50g / L and maltose 50g / L, and the solvent used to prepare the balance solution is 0.9% medical saline.

[0029] Fe 3 o 4 The average particle size of the nano magnetic beads is 1000nm.

Embodiment 2

[0030] The virus enrichment concentration method of embodiment 2 clinical samples

[0031] Take 2ml each of CVB3 and RSV virus liquid respectively (the virus titer is 10 5 PFU / ml), 2 mL of DMEM medium was used for experimental detection.

[0032] Detection process:

[0033] 1. Dissolve 100mg magnetic beads in 100μl balance solution and mix well;

[0034] 2. Carry out testing according to the following groups:

[0035] The first group (before enrichment): 1ml CVB3 virus solution and 100μl primary antibody mouse anti-CVB3 (1:500) were mixed upside down for 20min, then 100μl secondary antibody anti-mouse IgG (1:500) was added and mixed upside down for 20min;

[0036] The second group (experimental group): 1ml CVB3 virus solution and 100μl primary antibody anti-mouse anti-CVB3 (1:500) were mixed upside down for 20 minutes, then 100μl secondary antibody anti-mouse IgG (1:500) was added and mixed upside down for 20 minutes;

[0037] The third group (control group): 1ml DMEM and ...

Embodiment 3

[0048] Embodiment 3 equilibrium liquid stability test

[0049] 1. Thermal stability test

[0050] Place the reagent sample (the balance liquid in Example 1 and the nano-magnetic beads after the balance liquid treatment) in a water bath at 40°C for 3 days in a dark place, no delamination was found by visual observation, and no crystals were precipitated under the microscope, and then the test was carried out according to the above operation steps. The results showed that the enrichment effect was not weakened.

[0051] 2. Cold stability test

[0052]The reagent sample was placed in a -20°C refrigerator out of light for one month, during which it was repeatedly frozen and thawed several times, and then taken out, no stratification was found under the naked eye, and no crystals were precipitated under the microscope. weakened.

[0053] 3. Stability test at room temperature

[0054] The reagent sample was kept at room temperature and protected from light for 1 month. No delami...

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Abstract

The invention provides a virus biological magnetic bead enrichment and concentration kit and its application. The kit contains balance solution and Fe 3 o 4 nano magnetic beads. The composition of the balance liquid is as follows: glucose 100-200g / L, mannose 50-100g / L, galactose 20-50g / L and maltose 20-50g / L, prepared with physiological saline. Utilizing the kit of the present invention for virus enrichment and concentration can widely and efficiently adsorb viruses in clinical samples, such as enterovirus, respiratory syncytial virus, etc., and the kit has an enrichment effect on bacteria, is low in cost, and is easy to operate , without centrifugation, ensuring biological safety. The kit provided by the invention has the advantages of simple operation, rapidity, good specificity, high sensitivity, and low hardware requirements, and can be widely used in disease prevention and control at all levels and medical institutions.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a virus biological magnetic bead enrichment and concentration kit and its application. Background technique [0002] In recent years, due to the gradual increase in the number of tumor patients, organ transplantation, and immunodeficiency and immunocompromised patients, the incidence of related viral infections has greatly increased. Especially during the incubation period of virus infection, there is no characteristic symptom, so it is difficult to diagnose early clinically. For virus infection during the incubation period, the liquid samples collected in the body, such as tissue fluid, blood, cerebrospinal fluid, pleural and peritoneal fluid, urine, and sputum, have a small amount of virus. When the samples are directly tested by fluorescent quantitative PCR, due to the limitations of the detection method, It is difficult to detect a small amount of virus, and there are certain fa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/02
CPCC12N7/00
Inventor 韩俊王文军宋娟沈弢张浩王瑞芳刘宓史冰田宋芹芹梅国勇夏志强夏冬
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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