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CoxA16 virus strain and human CoxA16 inactivated vaccine

A technology of inactivated vaccines and virus strains, applied in antiviral agents, viruses/phages, biochemical equipment and methods, etc. Originality, high safety effect

Active Publication Date: 2012-12-26
JIANGSU CONVAC BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, although CoxA16 virus infection has caused more disease in various regions of my country, there is still no effective specific vaccine application

Method used

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  • CoxA16 virus strain and human CoxA16 inactivated vaccine
  • CoxA16 virus strain and human CoxA16 inactivated vaccine
  • CoxA16 virus strain and human CoxA16 inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Preparation of CoxA16 Inactivated Vaccine Virus Strain GX-20K-D

[0034] (1) The preparation method of CoxA16 inactivated vaccine virus strain GX-20K-D, comprising the following steps:

[0035] 1. The CoxA16 virus strain isolated from patients with hand, foot and mouth disease, as for the patient's throat swab and a small amount of herpes secretion, diluted with sterile PBS solution and filtered, inoculated in a single layer of KMB17 cells by suspension adsorption. 10 days after the lesion occurred, the KMB17 cells continued to be passaged for 4 passages. First, take 1ml of the virus liquid, perform RT-PCR according to the conventional method, obtain the cDNA sequence of the viral gene in segments, and connect the fragment into the puc-18 vector according to the conventional method, perform conventional nucleotide sequencing analysis, and use the sequence The results were confirmed by comparison in similar gene pools.

[0036] 2. The virus strain was ino...

Embodiment 2

[0047] Example 2 Preparation of CoxA16 Inactivated Vaccine

[0048] The preparation method of the CoxA16 inactivated vaccine of the present embodiment comprises the following steps:

[0049] 1. According to the KMB17 cell production and verification regulations, select the 28th generation KMB17 cells with dense growth, discard the culture medium, digest the cells with 0.1% trypsin, discard the trypsin solution, and use 4% of the volume of the original culture medium, DMEM Suspension cells.

[0050] 2. Add the 14th generation CoxA16 virus solution of the present invention to the above cell suspension, add 0.2-0.5ml of the virus solution, the moi is 0.02-0.1, mix well and incubate at 37°C for 30 minutes, and add light Shake, then transfer the suspension into a culture bottle, add an appropriate amount of cell growth medium, culture at 37°C for 3 to 4 days, and harvest after the cells adhere to the wall and complete lesions appear.

[0051] 3. Add formaldehyde to the virus ha...

Embodiment 3

[0063] Example 3 Safety Testing of CoxA16 Inactivated Vaccine Finished Products

[0064] The finished vaccine is injected into the peritoneal cavity of 18-20g mice with a weight of 0.5ml / mouse, a total of 20, and there should be no death within one month (routine abnormal toxicity test: 20 mice with a weight of 18-20g are injected with 0.5ml / mouse of the finished vaccine) Mice, fed for 7 days, without death, weight gain (about 1-10 grams); 2 guinea pigs, intraperitoneal injection of 5ml vaccine finished products, feeding for 7 days, no death, weight gain (about 1-40 grams) was judged as Qualified), the weight gain of the mice should be the same as that of the normal saline control group (see Table 2).

[0065]

[0066] The formaldehyde content of the finished vaccine should not be higher than 50μg / dose, and the bacterial endotoxin should not be higher than 100EU / dose. Should be negative in sterility testing (see Table 3).

[0067]

[0068]

[0069] Organization A...

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Abstract

The invention provides a CoxA16 virus strain, named CoxA16 (GX-20K-D virus strain) with CGMCCNo.6350 and including three bases, namely an original seed base, a master seed base and a working seed base; in addition, the CoxA16 virus strain has a nucleotide sequence shown as SEQNo.1. The invention has the following obvious beneficial effects: 1, a virus strain is provided for preparing a human CoxA16 inactivated vaccine and has effective immunogenicity and good growth and replication capacity on cells; 2, a production and preparation method of the human CoxA16 inactivated vaccine which is prepared from KMB17 human diploid cell matrix is provided, and the vaccine product has excellent immunogenicity and safety; and a suspended absorption technical method is provided so as to ensure the effective infection of the CoxA16 inactivated vaccine virus strain on KMB17 cell and excellent growth of the CoxA16 inactivated vaccine virus strain on the matrix.

Description

technical field [0001] The invention relates to a CoxA16 virus strain and an inactivated CoxA16 vaccine for humans, belonging to the field of biotechnology. Background technique [0002] Coxsackievirus A 16 Strain (CoxA16) is one of the main pathogens causing Hand, Foot and Mouth Disease (HFMD) in children. As a member of enteroviruses, CoxA16 has typical ribonucleic acid virus characteristics in structure, including its single-stranded positive-strand RNA genome, and the capsid protein structure composed of VP1, VP2, VP3, and VP4. After the virus infects the human body, it is usually transmitted through intestinal excretion→oral transmission, and in certain cases, it can also be transmitted through the respiratory tract. The infection caused by this virus is similar to Enterovirus 71 (EV71) infection, usually manifested as characteristic herpes on the hands, feet and mouth, accompanied by a certain degree of fever and cold symptoms. There are also a certain proportion of...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N7/02A61K39/125A61P31/14C12R1/93
Inventor 姜莉李琦涵董承红刘龙丁王晶晶谢忠平崔萍芳王丽春刘建生邵聪文廖芸赵树栋
Owner JIANGSU CONVAC BIO TECH CO LTD
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