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Human enterovirus 71-type C4-subtype lethal strain SD095 and its application

A human enterovirus, SD095 technology, applied in the field of virology and immunology, can solve the problems of low vaccine quality, inability to fully evaluate the vaccine effect by detection means, and inconspicuous clinical effect, and achieve obvious lethal effect and virulence Strong and fast lesion speed effect

Active Publication Date: 2014-06-04
SINOVAC BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The common strains of EV71 are all non-lethal strains, and only serological detection can be carried out in vitro. A single detection method cannot comprehensively evaluate the vaccine effect. The clinical effect of the antibody vaccine obtained from non-lethal strains as antigens is not obvious. relatively low quality vaccines

Method used

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  • Human enterovirus 71-type C4-subtype lethal strain SD095 and its application
  • Human enterovirus 71-type C4-subtype lethal strain SD095 and its application
  • Human enterovirus 71-type C4-subtype lethal strain SD095 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Acquisition of human enterovirus 71 C4 subtype lethal virus strain SD095

[0041] 1. Strain sample collection: Collect virus strain samples in the concentrated outbreak areas of HFMD across the country, obtain fecal or throat swab specimens aseptically according to the procedures specified in the "Guidelines for the Prevention and Control of Hand, Foot and Mouth Disease (2008 Edition)", and soak the specimens in physiological saline , and stored in -20℃ refrigerator.

[0042] Table 1 Sample information of C4 genotype EV71 strain

[0043]

[0044] 2 Isolation and passage of strains

[0045] The cells used for virus isolation are 129-generation African green monkey kidney cells (Vero) working seed batch (derived from ATCC 120-generation Vero cell seeds). for SH200500128). The source and passage history are clear, there is no exogenous contamination, and it is sensitive to EV71.

[0046] After the samples were processed, they were inoculated with Vero cell...

Embodiment 2

[0066] Example 2 Human enterovirus 71 C4 subtype lethal strain SD095 challenge protection experiment

[0067] In the present invention, the parental female mice are immunized intraperitoneally with different doses of enterovirus type 71 inactivated vaccine, and the suckling mice born to them are intraperitoneally challenged with a lethal dose of virus, and the survival rate of the suckling mice is observed to evaluate the protective effect of the vaccine, in order to determine the vaccine for human use. The immunization dose and immunization program provide the basis.

[0068] Cultivate Vero working cell seeds, inoculate EV71 virus seeds after culturing at 35±2°C for 2-5 days. After virus inoculation, inoculate the inoculated cells at 30±2°C for 10-15 days. According to the cytopathic condition, harvest the cell supernatant to obtain The virus harvest solution was inactivated by a formaldehyde solution with a final concentration of 180±30 μg / ml. Then, the inactivated solution...

Embodiment 3

[0070] Example 3 Establishment of animal lethal model based on C4 genotype EV71 virus lethal strain SD095

[0071] Detection of viral infection titer TCID of C4 genotype EV71 virus lethal strain by cytopathic assay 50 , then according to TCID 50 The test results were double-diluted, and intraperitoneal immunization was performed on suckling mice under 7 days of age, and the injection titers were 1TCID. 50 , 0.1TCID 50 , 0.01TCID 50 and 0.001TCID 50 , the injection dose is 50 μl. 10 suckling mice were immunized for each titer.

[0072] The mortality rate of suckling mice was counted after 7 days of immunization, and the results showed that 1 TCID 50 and 0.1TCID 50 All the suckling mice in the experimental group died within 7 days, and the lethality rate was 100%; 0.01TCID 50 and 0.001TCID 50 In the experimental group, 2 animals died within 7 days, and the fatality rate was 20%.

[0073]The above results show that when the virus titer is 0.1TCID 50 When immunizing suc...

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Abstract

The invention provides a human enterovirus71-type C4-subtype lethal strain SD095 with the accession number of CGMCC (China General Microbiological Culture Collection Center) NO.6601 and its application. A rat adaptive lethal strain is acquired by conventional virus isolation cultivation, animal infection and a series of technical means, and an animal lethal model based on the virus strain SD095 can be established. The acquisition of the lethal virus strain and the establishment of the animal lethal model have important significance for promoting clinical treatment and researches in China on diseases and death cases caused by EV71 (human enterovirus 71) virus, and have important values for further promotion of experimental and clinical studies and epidemiologic studies on the EV71, and new vaccine development and evaluation.

Description

technical field [0001] The invention belongs to the field of virology and immunology, and in particular relates to human enterovirus 71 C4 subtype lethal strain SD095 and its application. Background technique [0002] Hand, foot and mouth disease (HFMD) is a fecal-oral borne disease that is widespread worldwide and has been reported in most parts of the world. For a long time, mild hand, foot and mouth disease outbreaks periodically around the world. The disease was first reported in New Zealand in 1957, the coxsackie virus was first isolated in 1958, and the name HFMD was proposed in 1959. Enterovirus 71 (EV71) was first isolated from fecal specimens of infants with central nervous system diseases in the United States in 1969. The interval between HFMD epidemics is 2-3 years. The early pathogens are mainly CoxA16, and later EV71 and CoxA16 appear alternately, becoming the main pathogens of HFMD. In the mid-1970s, the EV71 epidemic with the central nervous system as the m...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/125A61K39/42A61P31/14C07K16/10G01N33/569C12R1/93
Inventor 薛晨宝郝超阴彦辉王琳高强
Owner SINOVAC BIOTECH
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