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《Chinese Journal of Veterinary Science》 2007-01
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Establishment and employment of PCR technique for distinguish Marek′s disease virus vaccine strain CVI988 from other MDV strains

DING Jia-bo1, JIANG Shi-jin1,ZHU Hong-fei2, CUI Zhi-zhong 1* (1.College of Animal Science and Technology, Shandong Agricultural University, Taian, Shandong 271018, China;2. Bioproduct Engineering Technology Center, Chinese Academy of Agricultural Sciences, Beijing 100081, China)  
The promoter of pp38 gene in Marek′s disease virus(MDV) vaccine strain CVI988 had a consecutive 5 bp(5′-AGCCG-3′)deletion. In this study, two pairs of primers were designed and synthesized to distinguish CVI988 strain from other MDV strains by polymerase chain reaction (PCR). 8 MDV strains (vaccine strains CVI988 and 814, virulent reference strains GA, very virulent reference strains RB1B, Md5 and Md11,very virulent plus reference strains 648A,and a Chinese field strain J-E) were distinguished by this technique and the detected result was consisted with the putative. Chicken tissue nucleic acid extracts from MDV-suspected chickens were used as the templats for PCR, and 7 samples out of 16 were MDV positive in the detection. At the same time, lymphocytes were isolated from the sixteen chickens for cell culture in chicken embryo fibroblast (CEF), respectively. In the second passage, only 7 samples of the PCR positive produced MDV specific plaque, the other 9 samples could not even when the cells were passed to the fifth passage. The cultivated cells were further detected by MDV specific monoclonal antibodies and confirmed the result of PCR. The above results indicated that the established PCR in this study was sensitive enough to distinguish MDV CVI988 strain from other MDV strains.
【Fund】: 国家自然科学基金重点资助项目(30300450);; 国家自然科学基金资助项目(30070544)
【CateGory Index】: S852.65
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