鸭坦布苏病毒js2010株的分离鉴定及间接elisa监测方法的建立-isolation and identification of duck tembusu virus js 2010 strain and establishment of indirect elisa monitoring method.docxVIP

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鸭坦布苏病毒js2010株的分离鉴定及间接elisa监测方法的建立-isolation and identification of duck tembusu virus js 2010 strain and establishment of indirect elisa monitoring method.docx

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鸭坦布苏病毒js2010株的分离鉴定及间接elisa监测方法的建立-isolation and identification of duck tembusu virus js 2010 strain and establishment of indirect elisa monitoring method

AbstractIn the spring of 2010, a severe disease was found in several duck farms in east-central China, major clinic characters included a sudden decline of feed uptake accompanied by a heavy drop in egg production, ataxia of gait, indeed, a few of infected egg-laying ducks stoped laying eggs or died. So far, the disease has spread to most of the duck-producing regions in China including Shanghai Autonomous City, Jiangsu Province, Anhui Province, Shandong Province and Hebei Province and caused serious economic loss to the duck industry in China. It is of great significance to determine the pathogen of the disease and to establish its rapid diagnostic methods.Firstly, to isolate and identify the pathogen that caused disease in ducks, the samples of diseased tissue of the infected ducks from a duck farm in Jiangsu Province were homogenized. Then the filtered homogenates were inoculated into several 10-day-old SPF chicken embryonated eggs (0.2mL/embryo) via the allantoic sac.The chick embryos died post-inoculation 4-6 days with the chick chorilallantoic membranes thickening andembryos swollen、hyperemia. The embryo allantoic fluid was collected and it was namedJS2010 strain which had an infectivity titer of 103.68ELD50/0.2mL.We make fortether studise on the disease by hemagglutination test, virus culture, histopathological examination and nucleic acid identification. The results showed that the isolated virus had no ability to agglutinate 1% erythrocytes including the chicken, duck, goose and swine. The virus could replicate in BHK-21 cells but not in CEF or Vero, the virus fluid had an infectivity titer of 105.33TCID50/0.2mL. The main expressions of CPE consisted in decreased cell diopter, rounding reduced cells, and cell exfoliation. The fatty degeneration of hepatocytes, intestinal bleeding and intestinal epithelial injury were found by pathologic observation. By RT-PCR and sequence analysis, the result showed that the isolated JS2010 strain s