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| Isolation Human Rotavirus Strains and Adaptive Culture |
| LI Song1, ZHANG Guang-ming1, WU Jin-yuan1, YIN Na1, YI Shan1, MI Kai1, CAO Ying2, LI Yang2, SUN Mao-sheng1, LI Hong-jun1 |
1. Institute of Medical Biology, Chinese Academy of Medical Science, Peking Union Medical College, Yunnan Key Laboratory of Vaccine Research & Development on Severe Infectious Diseases, Kunming 650018, China;
2. Kunming Medical University, Kunming 650031, China |
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Abstract Objective: To investigate separation method,the condition for culture and biological properties of wild rotavirus strain. Methods: To the qualitative detection of samples collected rotavirus use colloidal gold, PCR, PAGE.Process the positive samples according to the conventional method, and then do the separation of tissue culture. To analyze and evaluate the genome, gene sequences and proliferation kinetics of different transfer of culture samples. Using the positive serum of the G1P[8] rotavirus to do the virus neutralizing identification test. Results: By testing, the ZTR-68 rotavirus belong to A group were got, the arrangement of genome is 4:2:3:2,the genotype is G1P[8].The CPE was observed after the adapted culture the rotavirus,proliferated in MA104 cells, in the Vero cells. And the typical rotavirus form can be observed by using electron microscope. The processing is stable, which the strain proliferated in the Vero cells. And the infectious titer reached 7.25 log CCID50/ml, the peak time of proliferation is at 96h.There was not any other virus except the rotavirus in the culture medium of virus neutralizing identification test. Conclusion: The separated strain of human rotavirus, named ZTR-86,which was got from the separation of the diarrhea samples, has a good adaptability to the tissue culture and a good genetic stability. It provides a basis for the further study of its biological properties and the making of corresponding vaccine.
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Received: 06 November 2012
Published: 25 April 2013
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Cite this article:
LI Song, ZHANG Guang-ming, WU Jin-yuan, YIN Na, YI Shan, MI Kai, CAO Ying, LI Yang, SUN Mao-sheng, LI Hong-jun. Isolation Human Rotavirus Strains and Adaptive Culture. China Biotechnology, 2013, 33(4): 9-14.
URL:
https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2013/V33/I4/9
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