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首页> 外文期刊>Virus Genes >Genetic characterization of E2 gene of classical swine fever virus by restriction fragment length polymorphism and phylogenetic analysis.
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Genetic characterization of E2 gene of classical swine fever virus by restriction fragment length polymorphism and phylogenetic analysis.

机译：通过限制性片段长度多态性和系统发育分析对经典猪瘟病毒E2基因进行遗传鉴定。

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An RT-nested PCR (RT-nPCR)-based restriction fragment length polymorphism (RFLP) analyses of the E2 gene were developed for genetic subtyping and differentiation of vaccinated and infected classical swine fever virus (CSFV) strains. RT-nPCR identified 96 CSFV-positive samples from 321 clinical specimens from southeastern China during 2003-2008. The PCR products of positive samples were further differentiated using MspI digestion, 23 were identified as the C-strain, 62 as field strains, and 11 as mixture of the vaccine strain and field ones. RFLP with BglI, DdeI, DraI, and PstI were then used for subtyping of the field CSFV isolates. Thirty-eight field isolates phylogenetically classified as subgroup 2.1 based on E2 were divided into 11 subtypes by this RFLP scheme. Both RFLP profiling and sequence-based phylogenetic analysis revealed genetic diversity of CSFV in the field. Three novel substitutions at amino acid positions 17, 93, and 286 were identified in the predominant subtype VI strains isolated in 2008 as compared to other strains including historical subtype VI strains. These results suggest that CSFV in China experienced gradual variations and evolutionary accumulation progress. Thus, the RFLP methods targeting on the CSFV E2 gene are suitable for epidemiological survey in endemic area where the C-strain is applied for vaccination. Combination of the RFLP schemes with sequence-based phylogenetic analysis could provide more detailed information on transmission of CSFV in the region or even its evolution.
机译：E2基因的基于RT嵌套式PCR（RT-nPCR）的限制性片段长度多态性（RFLP）分析已开发用于疫苗接种和感染的经典猪瘟病毒（CSFV）菌株的遗传分型和分化。 RT-nPCR在2003-2008年期间从中国东南部的321个临床标本中鉴定出96个CSFV阳性样品。使用MspI消化进一步区分阳性样品的PCR产物，鉴定出23种为C株，鉴定为62种为田间菌株，11种为疫苗株和野外菌株的混合物。然后将具有BglI，DdeI，DraI和PstI的RFLP用于现场CSFV分离株的亚型分析。根据该RFLP方案，根据E2在系统发育上分类为2.1亚组的38个野外隔离株被分为11个亚型。 RFLP分析和基于序列的系统发育分析均揭示了该领域CSFV的遗传多样性。与其他菌株（包括历史性VI型菌株）相比，2008年分离出的主要VI型菌株中在氨基酸第17、93和286位上发现了三个新的取代。这些结果表明中国的猪瘟病毒经历了逐渐的变异和进化积累的进展。因此，针对CSFV E2基因的RFLP方法适用于在使用C株进行疫苗接种的流行地区的流行病学调查。 RFLP方案与基于序列的系统发育分析相结合可以提供有关该地区CSFV传播甚至进化的更详细信息。

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