Human rotavirus-A seed strain ZTR-68, and isolation, culture and identification thereof

A rotavirus and strain technology, applied in the directions of virus/phage, analytical materials, inactivation/attenuation, etc., can solve the problems of high cost of cold chain storage system, intussusception and virulence reversion mutation, etc. The effect of improving the efficiency of primary culture, high virus recovery yield, and high impurity removal effect

Active Publication Date: 2012-08-01
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the rotavirus vaccines in preclinical research or on the market are all oral attenuated live vaccines. Although the live attenuated vaccines have certain effects, they have defects in safety, such as causing intussusception and virulence regression mutations and producing new viruses. The risk of mutant strains; and the cold chain preservation system requires high cost. Therefore, the development of safe and effective inactivated rotavirus vaccines not only has advantages in avoiding the problems in the use of attenuated live vaccines, but also provides a basis for future development of combined vaccines. Provide the basis

Method used

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  • Human rotavirus-A seed strain ZTR-68, and isolation, culture and identification thereof
  • Human rotavirus-A seed strain ZTR-68, and isolation, culture and identification thereof
  • Human rotavirus-A seed strain ZTR-68, and isolation, culture and identification thereof

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Embodiment 1

[0033] Embodiment 1, the separation method of ZTR-68 strain involved in the present invention:

[0034] (1) Obtain virus samples

[0035] Collect diarrhea excrement samples from pediatric diarrhea patients in the hospital, resuspend in 0.01 M PBS, pH 7.2 solution, centrifuge at 12000 g for 20 minutes to get the supernatant, and use a 0.22 μm filter (purchased from Millipore) to sterilize and filter,- Store at 20°C for later use.

[0036] (2) Adaptation of virus on MA104 cells

[0037] Use 0.25% trypsin-EDTA to digest the P 29 generation MA104 cells that have formed a monolayer. 5.0 cells / ml. Press 2×10 5.0 Cells / bottle were inoculated into a small square bottle, and after culturing at 37°C for 3 to 4 days, inoculated 200 μl of the diluent of diarrhea samples from rotavirus-infected children on the cell monolayer, adsorbed at 37°C for 2 hours, and then added calf serum-free DMEM cell maintenance solution, cultivated at 37°C and observed cell changes. After 3 to 4 days of cy...

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Abstract

A human rotavirus-A seed strain ZTR-68 is submitted to CGMCC (china general microbiological culture collection center) for preservation in September 22th, 2011, and encoded as CGMCC No.5265. The strain is a human rotavirus wide-type single purified strain, which is high in virus replication and fine in genetic stability. The human rotavirus-A seed strain ZTR-68 is applicable to developing and producing virus seeds for human rotavirus vaccines, and applicable to attenuated rotavirus oral vaccines and inactivated human rotavirus injection vaccines. The invention further provides a method for isolation, culture and identification of the human rotavirus-A seed strain ZTR-68.

Description

technical field [0001] The invention relates to the separation and identification of rotavirus, especially human rotavirus seed strain and the immunological effect evaluation of the strain. Background technique [0002] Human rotavirus is one of the most important pathogens causing diarrhea in infants and young children worldwide. Among infants under the age of 5 in the world, there are 17 million cases of acute gastroenteritis caused by rotavirus infection every year, causing about 40,000 infant deaths, and 80% of these deaths occur in low-income countries in Asia and Africa. income in developing countries. In my country, the positive rate of rotavirus detection among infants under 5 years old hospitalized due to diarrhea is about 50%. Rotavirus infection in infants and young children mainly manifests as sudden onset, watery stools, vomiting, fever and dehydration, and eventually severe dehydration and electrolyte imbalance lead to death in severely ill infants and young ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/08G01N33/569C12Q1/70C12Q1/68C12R1/93
Inventor 李泓钧孙茂盛易山张光明吴晋元
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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