Germain Karine, Valas Stephen
Agence Française de Sécurité Sanitaire des Aliments, Laboratoire d'Etudes et de Recherches Caprines, B.P. 3081, 60 rue de Pied de Fond, F-79012 Niort Cedex, France.
Virus Res. 2006 Sep;120(1-2):156-62. doi: 10.1016/j.virusres.2006.03.002. Epub 2006 Apr 17.
Small ruminants lentiviruses (SRLV) nucleotide sequences spanning the V1V2 variable regions of the env gene were amplified by nested-PCR from 38 blood samples collected from 16 naturally infected sheep flocks in France. For the rapid SRLV group determination of field isolates, the PCR-amplified fragments were subjected to a SRLV-adapted heteroduplex mobility assay (HMA). All viral sequences were clearly assignable to the SRLV group B by HMA analysis. Twenty-seven SRLV isolates were selected for DNA sequence analysis. In each case, nucleotide comparison and phylogenetic analyses confirmed the genetic relationships inferred by HMA. Six SRLV isolates belonged to subtype B1, and 21 pertained to subtype B2, one flock being infected with both subtypes. Subtypes B1 and B2 were found with different frequencies and geographic spread, but exhibited similar genetic diversities. These results give a more complete picture of the distribution and heterogeneity of SRLV env subtypes in sheep and confirmed that multiple interspecies transmission occurred in the past. Furthermore, HMA appeared to be a rapid and reliable method to differentiate caprine arthritis encephalitis virus from maedi-visna virus.
通过巢式聚合酶链反应(nested-PCR),从法国16个自然感染绵羊群采集的38份血液样本中扩增出小反刍兽慢病毒(SRLV)包膜基因V1V2可变区的核苷酸序列。为了对野外分离株进行快速的SRLV分组测定,将聚合酶链反应扩增片段进行SRLV适应性异源双链迁移率分析(HMA)。通过HMA分析,所有病毒序列均可明确归为SRLV B组。选择27株SRLV分离株进行DNA序列分析。在每种情况下,核苷酸比较和系统发育分析均证实了HMA推断的遗传关系。6株SRLV分离株属于B1亚型,21株属于B2亚型,一个羊群同时感染了这两种亚型。发现B1和B2亚型出现频率和地理分布不同,但遗传多样性相似。这些结果更全面地展现了SRLV env亚型在绵羊中的分布和异质性,并证实过去发生了多次种间传播。此外,HMA似乎是一种区分山羊关节炎脑炎病毒和梅迪-维斯纳病毒的快速可靠方法。