Chua K B, Wang L F, Lam S K, Crameri G, Yu M, Wise T, Boyle D, Hyatt A D, Eaton B T
Department of Medical Microbiology, University of Malaya Medical Center, 50603 Kuala Lumpur, Malaysia.
Virology. 2001 May 10;283(2):215-29. doi: 10.1006/viro.2000.0882.
A search for the natural host of Nipah virus has led to the isolation of a previously unknown member of the family Paramyxoviridae. Tioman virus (TiV) was isolated from the urine of fruit bats (Pteropus hypomelanus) found on the island of the same name off the eastern coast of peninsular Malaysia. An electron microscopic study of TiV-infected cells revealed spherical and pleomorphic-enveloped viral particles (100--500 nm in size) with a single fringe of embedded peplomers. Virus morphogenesis occurred at the plasma membrane of infected cells and morphological features of negative-stained ribonucleoprotein complexes were compatible with that of viruses in the family Paramyxoviridae. Serological studies revealed no cross-reactivity with antibodies against a number of known Paramyxoviridae members except for the newly described Menangle virus (MenV), isolated in Australia in 1997. Failure of PCR amplification using MenV-specific primers suggested that this new virus is related to but different from MenV. For molecular characterization of the virus, a cDNA subtraction strategy was employed to isolate virus-specific cDNA from virus-infected cells. Complete gene sequences for the nucleocapsid protein (N) and phosphoprotein (P/V) have been determined and recombinant N and V proteins produced in baculovirus. The recombinant N and V proteins reacted with porcine anti-MenV sera in Western blot, confirming the serological cross-reactivity observed during initial virus characterization. The lack of a C protein-coding region in the P/V gene, the creation of P mRNA by insertion of 2-G residues, and the results of phylogenetic analyses all indicated that TiV is a novel member of the genus Rubulavirus.
对尼帕病毒自然宿主的搜寻导致了副粘病毒科一个此前未知成员的分离。从在马来西亚半岛东海岸同名岛屿上发现的果蝠(淡黄果蝠)尿液中分离出了刁曼病毒(TiV)。对感染TiV的细胞进行的电子显微镜研究显示,病毒颗粒呈球形和多形性包膜(大小为100 - 500纳米),有一圈嵌入的纤突。病毒形态发生在受感染细胞的质膜上,负染核糖核蛋白复合物的形态特征与副粘病毒科病毒相符。血清学研究表明,除了1997年在澳大利亚分离出的新描述的门果病毒(MenV)外,该病毒与针对多种已知副粘病毒科成员的抗体无交叉反应。使用MenV特异性引物进行PCR扩增失败,表明这种新病毒与MenV相关但不同。为了对该病毒进行分子特征分析,采用了cDNA消减策略从病毒感染细胞中分离病毒特异性cDNA。已经确定了核衣壳蛋白(N)和磷蛋白(P/V)的完整基因序列,并在杆状病毒中产生了重组N和V蛋白。重组N和V蛋白在蛋白质印迹中与猪抗MenV血清发生反应,证实了在最初病毒特征分析期间观察到的血清学交叉反应。P/V基因中缺乏C蛋白编码区、通过插入2个G残基产生P mRNA以及系统发育分析结果均表明TiV是风疹病毒属的一个新成员。