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猪圆环病毒2型陕西杨凌株的分离鉴定及全基因组分析
汤智慧1, 郭抗抗1, 张彦明1
西北农林科技大学 动物医学院
摘要:
【目的】 分离得到猪圆环病毒2型(PCV-2)陕西杨凌株,测定病毒滴度(TCID50),并进行全基因组序列比对,为研制针对性更强的PCV-2疫苗提供理论依据。【方法】 从经PCR检测为猪圆环病毒2型阳性的病料中,分离病毒并接种PK-15细胞,通过PCR检测、间接免疫荧光试验及电镜观察,对分离病毒进行鉴定;用免疫过氧化物酶细胞单层试验(IPMA),测定分离毒株的TCID50,并对其全基因组序列进行测定与分析。【结果】 分离得到1株猪圆环病毒2型毒株,分离株在PK-15细胞上的TCID50为10-4.75。全基因组序列同源性比对发现,分离株与一加拿大分离株(DQ200735)同源性最高(99.4%)。【结论】 从陕西杨凌分离得到了1个猪圆环病毒2型分离株。
关键词:  猪圆环病毒2型  分离鉴定  TCID50  陕西杨凌株
DOI:
分类号:
基金项目:陕西省“13115”科技创新工程重大科技专项(2007ZDKG-13)
Isolation and identification of porcine circovirus type 2 Yangling isolate and sequence analysis of the whole genomes
Abstract:
【Objective】The aim of the study was to isolate the porcine circovirus type 2(PCV-2) in Yangling and to determine the 50% tissue culture infective dose(TCID50)of the virus isolate,and to analyze the whole genes,which will make a foundation for vaccines developing.【Method】The PCV-2 was isolated from the PCV-2 positive blood samples detected by conventional PCR.The virus was isolated and identified with cell isolation and culture,PCR,indirect immunofluorescent assay (IFA) and electron microscopic observation.The complete genomes of the isolated strains were sequenced and analyed.The 50% tissue culture infective dose of isolate was determined with immunoperoxidase monolayer assay (IPMA).【Result】A PCV-2 Yangling strain was isolated from blood samples.The TCID50 of the isolate was 10-4.75.The homology of PCV-2 isolate with a PCV-2 Canadia isolate (DQ200735) was up to 99.4%.【Conclusion】A PCV-2 Yangling strain was isolated and some studies were carried on this isolate.
Key words:  porcine circovirus type 2  isolation and identification  50% tissue culture infective dose  Yangling isolate