Genotyping field strains of African swine fever virus by partial p72 gene characterisation
Genotyping field strains of African swine fever virus by partial p72 gene characterisation
复制标题
DOI：
10.1007/s00705-002-0946-8
复制
发表时间：
2003-04-01
影响因子：
2.7
通讯作者：
Thomson, GR
中科院分区：
文献类型：
--
作者：
Bastos, ADS;Penrith, ML;Thomson, GR
开发了一种基于PCR的测序方法，该方法允许在收到临床标本后5小时和48小时内分别检测和表征非洲猪瘟病毒（ASFV）变体。对应于p72基因C-末端的478 bp片段的扩增证实了病毒的存在，通过核苷酸序列测定和系统发育分析实现了遗传表征。该方法被应用到55个病毒，包括那些代表的主要ASF谱系以前确定的限制性片段长度多态性（RFLP）分析。结果证实，p72基因分型方法鉴定了相同的主要病毒群。使用基于PCR的方法对不同地理、物种和时间来源的其他病毒进行表征，表明非洲大陆存在10种主要的ASF基因型，其中最大的一种包括一组从欧洲、南美洲、加勒比和西非暴发中回收的遗传同质病毒（ESAC-WA基因型）。相比之下，来自南部和东部非洲国家的病毒是异质性的，在各个国家内存在多种基因型。本研究提供了一种快速、准确地确定ASF田间和暴发菌株基因型的方法，因此对ASF的分子流行病学澄清是有用的。
A PCR-based sequencing method was developed which permits detection and characterization of African swine fever virus (ASFV) variants within 5 and 48 h, respectively, of receipt of a clinical specimen. Amplification of a 478 bp fragment corresponding to the C-terminal end of the p72 gene, confirms virus presence with genetic characterization being achieved by nucleotide sequence determination and phylogenetic analysis. The method was applied to 55 viruses including those representative of the major ASF lineages identified previously by restriction fragment length polymorphism (RFLP) analysis. Results confirmed that the p72 genotyping method identifies the same major viral groupings. Characterization of additional viruses of diverse geographical, species and temporal origin using the PCR-based method indicated the presence of ten major ASF genotypes on the African continent, the largest of which comprised a group of genetically homogeneous viruses recovered from outbreaks in Europe, South America, the Caribbean and West Africa (the ESAC-WA genotype). In contrast, viruses from southern and East African countries were heterogeneous, with multiple genotypes being present within individual countries. This study provides a rapid and accurate means of determining the genotype of field and outbreak strains of ASF and is therefore useful for molecular epidemiological clarification of ASF.