Method for purifying porcine reproductive and respiratory syndrome virus
A technology of respiratory syndrome and purification method, applied in the field of porcine reproduction and respiratory syndrome virus purification, can solve the problems of poor purification effect, high cost, complicated operation of virus purification method, etc., so as to increase virus recovery efficiency and reduce purification Steps, the effect of meeting the needs of scientific research
Inactive Publication Date: 2018-06-29
扬州大学镇江高新技术研究院
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[0007] The present invention aims to provide an efficient and simple purification method of porcine reproductive and respiratory syndrome virus by combining PEG precipitation and dialysis purification, which has solved the problems of cumbersome operation, high cost and poor purification effect of the existing virus purification method
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[0026] 1. Materials and equipment
[0027] 1) The PRRSV commercialized vaccine strain JXA1-R strain and Marc-145 cell line were obtained from the Laboratory of Innate Immune Signaling, Immunity and Infection, School of Veterinary Medicine, Yangzhou University, Jiangsu Province.
[0028] 2) IMDM medium and fetal bovine serum were purchased from Gibco
[0029] 3) PEG6000 was purchased from MERCK, Germany, and the dialysis bag was purchased from Spectrum
[0030] 4) PBS (10X) was purchased from Jiangsu Biyuntian Biotechnology Co., Ltd.
[0031] 5) The refrigerated centrifuge was purchased from eppendorf, the ultra-clean bench, CO2 cell incubator, and HulaMixer were purchased from ThermoFisher
[0032] 2. Experimental method
[0033] 1) Virus propagation and preparation:
[0034] 1. Place the frozen Marc-145 cells in a water bath at 37°C for rapid thawing and put them into a cell bottle, and add 6 mL of IMDM medium with 10% fetal bovine serum (preheated at 37°C for 1 hour), an...
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The invention discloses a method for purifying porcine reproductive and respiratory syndrome virus. The method comprises the steps of virus centrifugation preliminary purification, PEG6000 precipitation method secondary purification, dialysis bag dialysis purification and the like. The method optimizes the concentration, precipitation time and centrifugal force of PEG6000 to maximize a recovery rate of virus. The porcine reproductive and respiratory syndrome virus purified by the method can meet the basic laboratory research and vaccine preparation requirements. Compared with the prior art, the product prepared by the invention has the advantages of high virus titer, good integrity and clear background, and the process is simple and the cost is low, so the method has a good promotion prospect.
Description
technical field [0001] The invention relates to a method for purifying virus, in particular to a method for purifying porcine reproductive and respiratory syndrome virus. Background technique [0002] Porcine reproductive and respiratory syndrome, also known as porcine blue ear disease, is a serious viral infectious disease in the pig industry at present, causing severe reproductive disorders and respiratory diseases in pigs. The virus belongs to the Arteriviridae family and is a non-segmented single-stranded positive-sense RNA virus. The disease first broke out in the United States in 1987 and has spread to almost all pig-raising countries, causing serious damage to the global pig industry. Huge loss. In 2006, highly pathogenic porcine reproductive and respiratory syndrome characterized by high fever and high mortality occurred in my country. The World Organization for Animal Health has listed porcine reproductive and respiratory syndrome as one of the animal diseases that...
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IPC IPC(8): C12N7/02C12R1/93
CPCC12N7/00C12N2770/10051
Inventor 周昕袁嘉晟
Owner 扬州大学镇江高新技术研究院