Separation method of severe fever with thrombocytopenia syndrome virus strain

A separation method and a syndrome technology, which are applied in the field of medical microbiology, can solve problems such as difficult cultivation, difficulty in isolating virus strains of fever with thrombocytopenia syndrome, and low purity of strains, achieving high purity, simple and easy operation, and low consumption. short-term effect

Pending Publication Date: 2021-09-03
李家斌
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the difficulties in the separation of fever with thrombocytopenia syndrome virus strains in the prior art, the isolated bacterial strains have low purity, and it is difficult to cultivate them in cells. A method for isolating virus strains of fever with thrombocytopenia syndrome

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] A method for isolating fever with thrombocytopenia syndrome virus strain proposed by the present invention comprises the following steps:

[0017] (1) Infect Vero cells (African green monkey kidney cells) with febrile thrombocytopenia syndrome virus and culture them for 5 days. After more than 60% of the cells have cytopathic effects, collect the cell culture supernatant;

[0018] (2) Use a 0.22 μm filter head to filter the cell culture supernatant collected in step (1) in a biological safety cabinet;

[0019] (3) Prepare an ultracentrifuge tube, add 8ml of 20% sucrose solution at the bottom of the ultracentrifuge tube to form a sucrose cushion;

[0020] (4) Along the wall of the ultracentrifuge tube in step (3), slowly add 30 ml of the cell culture supernatant obtained by filtering in step (2) on the sucrose cushion;

[0021] (5) Centrifuge the ultracentrifuge tube in step (4) at a temperature of 4°C, set the centrifugal speed to 40000g, and the centrifugation time to...

Embodiment 2

[0024] A method for isolating fever with thrombocytopenia syndrome virus strain proposed by the present invention comprises the following steps:

[0025] (1) Infect Vero cells (African green monkey kidney cells) with febrile thrombocytopenia syndrome virus, culture for 6 days, and collect the cell culture supernatant after more than 70% of the cells have cytopathic effects;

[0026] (2) Use a 0.22 μm filter head to filter the cell culture supernatant collected in step (1) in a biological safety cabinet;

[0027] (3) Prepare an ultracentrifuge tube, add 8ml of 20% sucrose solution at the bottom of the ultracentrifuge tube to form a sucrose cushion;

[0028] (4) Along the wall of the ultracentrifuge tube in step (3), slowly add 30 ml of the cell culture supernatant obtained by filtering in step (2) on the sucrose cushion;

[0029] (5) Centrifuge the ultracentrifuge tube in step (4) at a temperature of 4°C, set the centrifugal speed to 40000g, and the centrifugation time to 4h; ...

Embodiment 3

[0032] A method for isolating fever with thrombocytopenia syndrome virus strain proposed by the present invention comprises the following steps:

[0033] (1) Infect Vero cells (African green monkey kidney cells) with febrile thrombocytopenia syndrome virus and culture them for 7 days. After more than 80% of the cells have cytopathic effects, collect the cell culture supernatant;

[0034] (2) Use a 0.22 μm filter head to filter the cell culture supernatant collected in step (1) in a biological safety cabinet;

[0035] (3) Prepare an ultracentrifuge tube, add 8ml of 20% sucrose solution at the bottom of the ultracentrifuge tube to form a sucrose cushion;

[0036] (4) Along the wall of the ultracentrifuge tube in step (3), slowly add 30 ml of the cell culture supernatant obtained by filtering in step (2) on the sucrose cushion;

[0037] (5) Centrifuge the ultracentrifuge tube in step (4) at a temperature of 4°C, set the centrifugal speed to 40000g, and the centrifugation time to...

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Abstract

The invention belongs to the technical field of medical microbiology, particularly relates to a separation method of a severe fever with thrombocytopenia syndrome virus strain, and solves the problems that in the prior art, the severe fever with thrombocytopenia syndrome virus strain is difficult to separate, the strain obtained though separation is low in purity and is difficult to culture in cells, and no optimum scheme for separating the severe fever with thrombocytopenia syndrome virus strain exists in the prior art. The separation method of the severe fever with thrombocytopenia syndrome virus strain comprises the following steps of infecting Vero cells with the severe fever with thrombocytopenia syndrome virus, performing culturing, and collecting cell culture supernate; and performing filtration, adding a sucrose cushion layer, performing centrifugation, and taking a centrifugal precipitate to obtain a finished product. The separation method is simple, convenient and easy to operate, and consumes short time; and the severe fever with thrombocytopenia syndrome virus strain obtained through separation has high purity, can be used for subsequent research on pathogenic mechanisms and specific clinical manifestations of the severe fever with the thrombocytopenia syndrome, and has profound significance.

Description

technical field [0001] The invention relates to the technical field of medical microbiology, in particular to a method for isolating fever with thrombocytopenia syndrome virus strains. Background technique [0002] Severe fever with thrombocytopenia syndrome (SFTS) is an acute infectious disease caused by infection with bunyavirus (severe fever with thrombocytopenia syndrome bunyavirus, SFTSV), a new virus belonging to the family Bunyaviridae. The genus Virus was first discovered in China in 2009. In recent years, it has spread in more than half of China, causing thousands of people to get sick or even die. reports. The main clinical symptoms of the disease are high fever, anorexia, myalgia, chills, enlarged lymph nodes, leukopenia, thrombocytopenia, and multiple organ failure, etc. A small number of patients are seriously ill and develop rapidly, and may die due to multiple organ failure . According to the monitoring and investigation results by the end of 2011, most of ...

Claims

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Application Information

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IPC IPC(8): C12N7/00
CPCC12N7/00C12N2760/12251
Inventor 李家斌殷俊温萌胡立芬叶英刘艳艳程君陈振柳燕马雪娇卞婷婷
Owner 李家斌
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