Immunofluorescence method for detecting fever with thrombocytopenia syndrome virus infection IgM antibody and detection kit thereof

A virus infection and antibody detection technology, applied in the field of immunofluorescence detection, can solve the problems of high requirements for experimental instruments, unsuitable for rapid detection in primary hospitals and on-site, and cumbersome operation, and achieves low detection cost, low cost and high detection efficiency. Effect

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

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Problems solved by technology

[0003] The purpose of the present invention is to solve the problem that the detection method in the prior art has high requirements for experimental instruments and cumbersome operation. After the amplification is completed, the products need to be discriminated through steps such as electrophoresis and development, which are not suitable for basic hospitals and on-site rapid detection. , and proposed a fever with thrombocytopenia syndrome virus infection IgM antibody detection immunofluorescence method and its detection kit

Method used

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  • Immunofluorescence method for detecting fever with thrombocytopenia syndrome virus infection IgM antibody and detection kit thereof
  • Immunofluorescence method for detecting fever with thrombocytopenia syndrome virus infection IgM antibody and detection kit thereof
  • Immunofluorescence method for detecting fever with thrombocytopenia syndrome virus infection IgM antibody and detection kit thereof

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Effect test

Embodiment 1

[0031] A kind of fever with thrombocytopenia syndrome virus infection IgM antibody detection immunofluorescence method proposed by the present invention comprises the following steps:

[0032] (1) Infect Vero cells with fever and thrombocytopenia syndrome virus, culture for 5 days, and after more than 60% of the cells have cytopathic effect, collect the cell culture supernatant, and digest the cells with 0.25% trypsin;

[0033] (2) After completing the digestion operation in step (1), blow off the cells with DMEM liquid containing 3% serum;

[0034] (3) Collect the cells that have been blown off in step (2) and centrifuge in the ep tube, set the centrifugation speed to 300g, and the centrifugation time to 15min, and collect the precipitate after centrifugation;

[0035] (4) washing the precipitate collected in step (3) with sterile PBS twice;

[0036] (5) Regulate the cell concentration with PBS under a microscope while observing the precipitate that has been cleaned in step ...

Embodiment 2

[0047] A kind of fever with thrombocytopenia syndrome virus infection IgM antibody detection immunofluorescence method proposed by the present invention comprises the following steps:

[0048] (1) Infect Vero cells with Fever with Thrombocytopenia Syndrome Virus, culture for 6 days, after more than 70% of the cells have cytopathic effect, collect the cell culture supernatant, and digest the cells with 0.25% trypsin;

[0049] (2) After completing the digestion operation in step (1), blow off the cells with DMEM liquid containing 3% serum;

[0050] (3) Collect the cells that have been blown off in step (2) and centrifuge in the ep tube, set the centrifugation speed to 300g, and the centrifugation time to 15min, and collect the precipitate after centrifugation;

[0051] (4) washing the precipitate collected in step (3) with sterile PBS for 3 times;

[0052] (5) Regulate the cell concentration with PBS under a microscope while observing the precipitate that has been cleaned in st...

Embodiment 3

[0063] A kind of fever with thrombocytopenia syndrome virus infection IgM antibody detection immunofluorescence method proposed by the present invention comprises the following steps:

[0064] (1) Infect Vero cells with fever and thrombocytopenia syndrome virus, culture for 7 days, after more than 80% of the cells have cytopathic effect, collect the cell culture supernatant, and digest the cells with 0.25% trypsin;

[0065] (2) After completing the digestion operation in step (1), blow off the cells with DMEM liquid containing 3% serum;

[0066] (3) Collect the cells that have been blown off in step (2) and centrifuge in the ep tube, set the centrifugation speed to 300g, and the centrifugation time to 15min, and collect the precipitate after centrifugation;

[0067] (4) washing the precipitate collected in step (3) with sterile PBS twice;

[0068] (5) Regulate the cell concentration with PBS under a microscope while observing the precipitate that has been cleaned in step (4) ...

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Abstract

The invention belongs to the technical field of immunofluorescence method detection, particularly relates to an immunofluorescence method for detecting a fever with thrombocytopenia syndrome virus infected IgM antibody and a detection kit thereof, and solves the problems that in the prior art, the detection method has high requirements on experimental instruments, the operation is complicated, and the detection cost is high, and the prior art is not suitable for primary hospitals and on-site rapid detection due to the fact that products needs to be distinguished through electrophoresis, development and other steps after amplification is completed. The detection kit is composed of 12 holes, 2 microliters of cell suspension is dropwise added into each hole for hole laying and slide climbing, and after the cell suspension in the holes volatilizes, cold acetone is used for fixing; and serum of a patient is diluted with PBS and then added into antigen sheet holes, each antigen sheet is provided with a hole for blank control, and meanwhile positive control and negative control are set. The method has the advantages of relatively high specificity and sensitivity for detecting pathogens of the severe fever with thrombocytopenia syndrome, high detection accuracy, high detection efficiency and low detection cost.

Description

technical field [0001] The invention relates to the technical field of immunofluorescence detection, in particular to an immunofluorescence method for detecting IgM antibody of fever with thrombocytopenia syndrome virus infection and a detection kit thereof. 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 . Accordin...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N21/64
CPCG01N33/6854G01N33/56983G01N21/6486G01N21/6458G01N2333/175G01N2469/20
Inventor 李家斌殷俊温萌陈振刘艳艳叶英程君胡立芬柳燕马雪娇常啸卞婷婷
Owner 李家斌
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