Gould A R, Kattenbelt J A, Lenghaus C, Morrissy C, Chamberlain T, Collins B J, Westbury H A
CSIRO, Australian Animal Health Laboratory, Geelong Victoria, Australia.
Virus Res. 1997 Jan;47(1):7-17. doi: 10.1016/s0168-1702(96)01399-8.
The complete nucleotide sequence of the Czech strain of rabbit haemorrhagic disease virus (RHDV) was determined to be 7437 nucleotides in length with a 5-terminal non-coding region of 9 nucleotides and a 3'-terminal non-coding region of 59 nucleotides. Two open reading frames (ORFs) were found within this sequence coding for polypeptides of 2344 (nucleotides 10-7044) and 117 amino acids (nucleotides 7025-7378). The sequence of this isolate was approximately 1% different from that reported by Meyers et al., having 78 nucleotide changes which resulted in 30 amino acid differences, the majority of these clustering in the N-terminus of the large ORF and the middle of the viral coat protein. Only a single conservative amino acid change was seen in the smaller 3'-terminal ORF. Since the virus cannot at present be propagated in tissue culture, but isolated only after replication in rabbits, the reported sequence must be considered as a consensus sequence from the viral population. To gain some understanding of the possible sequence diversity within this virus population, 97 clones were sequenced from a polymerase chain reaction (PCR) fragment to determine the sequence diversity of the virus population. Four major classes of variant were described with mutations generally in the third base position of codons. A nested reverse transcriptase (RT) PCR (using sequence derived for the coat protein of RHDV) was used to determine the presence or absence of RHDV inoculated into non-host animal species. No replication of the virus was detected in 28 different vertebrate species other than rabbits. PCR tests on both mosquitoes and fleas feeding on RHDV infected rabbits were positive. The RT-PCR test was more sensitive when compared with an antigen capture ELISA to detect the presence of genomic RNA/or virus in infected rabbits.
已确定捷克兔出血病病毒（RHDV）毒株的完整核苷酸序列长度为7437个核苷酸，5'端非编码区有9个核苷酸，3'端非编码区有59个核苷酸。在该序列中发现了两个开放阅读框（ORF），分别编码2344个氨基酸（核苷酸10 - 7044）和117个氨基酸（核苷酸7025 - 7378）的多肽。该分离株的序列与迈耶斯等人报告的序列约有1%的差异，有78个核苷酸变化，导致30个氨基酸差异，其中大多数集中在大ORF的N端和病毒衣壳蛋白的中部。在较小的3'端ORF中仅观察到一个保守的氨基酸变化。由于该病毒目前无法在组织培养中增殖，只能在兔体内复制后分离得到，因此所报告的序列必须被视为病毒群体的一致序列。为了对该病毒群体内可能的序列多样性有所了解，对聚合酶链反应（PCR）片段的97个克隆进行了测序，以确定病毒群体的序列多样性。描述了四种主要的变异类型，突变通常发生在密码子的第三位。使用巢式逆转录酶（RT）PCR（使用从RHDV衣壳蛋白推导的序列）来确定接种到非宿主动物物种中的RHDV是否存在。除兔子外，在28种不同的脊椎动物物种中未检测到病毒复制。对吸食感染RHDV兔子血液的蚊子和跳蚤进行的PCR检测均呈阳性。与抗原捕获ELISA相比，RT-PCR检测在检测感染兔子中基因组RNA/或病毒的存在时更敏感。
