| HFRS is a natural disease which is spread by the mouse carrying the Hantaan virus. It is one of the most dangerous EHF in the world. In our country, the HAN type and SEO type is spreading every year which is the most harmful disease except for various heptis. It was discovered in 1930's in most countries .It was identified as EHF in 1942. It was renamed as HFRS by health administration in 1994.About 67 animals can be affected by Hanta virus in our country. Among there, there are Apodemus and Mus and big white mouse. Birds, house-fed animals such as cats, pigs can also amplify the Hanta virus's spreading. There are three spreading routes: animal, mite and vertical spreading.HFRS virus can attack various organizations and be harmful to whole body. The factual disease has three main points: fever, bleeding and injure to kidney .The prominent appearance is "three red", that is, the blood conflud and turn red in the patient's face, neck and front breast. Besides these, there are "three pain", that is, eye ache, headache and kidney ache and will occur five periods, that is, fever period, low blood pressure period, lack of urine period, full of urine period and recovery period. Besides this main appearance, there are a variety of hetero-appearances, such as lung bleeding, inflammatory brain membrane.There are 1.3 million diseases in 1950-1995 in the world .By the end of 2000,there are 1.62 million disease in the world. China has 1.45 million which is 90% of the whole world .The dead rate is 2.02%. Because the patients who have the virus will almost lose their heavy labor ability, it is very harmful to our country, especially in countryside.Vaccine is the most effective method preventing the disease .The vaccine study was started in late 1970's .Three vaccines were successfully manufactured in 1993: MGKC type I vaccine, GHKC type â…¡ vaccine and PSMB vaccine. The above three vaccines is prepared by adding the virus into the primary cells, after incubation lysis the cell and free the virus into medium, depleting the cell debris, adding inactivetor(BPL or formalin) and A1(OH)3.The two covalence vaccine is then prepared which includes type I and type I virus. Because the vaccine production use primary cell, there are a few shortages: 1.The large-scale production was restricted by we use of primary cell which use a lot of heath animals.2.In the period of production, it's hardly to avoid preventing exogenous factor into product.3.The antigen was unpurified and would arise serious side effect. The effective antigen is low and will arise failure of vaccine.In order to solve the problems in primary cell vaccine, it is very necessary to study Vero cell purified vaccine. We conduct a project to study the two covalence HFRS purified vaccine (Vero cell)'s middle scale production arts.1.Hemorrhagic fever with renal syndrome virus SD9805 (Hantavirus type 1) and HB9908 (Hantavirus type 2) were passed through three generations to obtain the toxicity about 7.0 logTCID50/ml. The two cell lines with high toxicity were passed through five and six generations to get toxicity about 7.0 logTCID50/ml. After 11 passages of SD9805 and 10 passages of HB9908, the toxicity reached their highest level (7.66 logTCID50/ml). The two cell lines were conducted neutralization test with typical virus cell lines, reaction and identification tests with different specific monoclonal antibodies. The results showed that SD9805 is Hantavirus type I and HB9908 is Hantavirus type â…¡. 2. Two kinds of molecular sieves media(Sepharose 4FF and Sephacryl 300HR)and two kinds of ion exchange media(HiTrapTM-DEAE Fast Flow and HiTrapTM-SP Fast Flow)were tested for effectiveness in vaccine purification. Considering the antigen amount and the result of SDS-PAGE analysis, we defined a production arts based on Sepharose 4FF. 3. The products manufactured according to the production arts were conducted three kinds of tests including abnormal toxity test, antigenity test, immunocity test and alergicity test. The results show... |
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