Santos P R, Azevedo M L B, Borges M B J, Freire M S, Nascimento J P, Moraes M T B
Bio-Manguinhos, Fundaço Oswaldo Cruz, Rio de Janeiro, RJ, Brasil.
Braz J Med Biol Res. 2003 Nov;36(11):1475-84. doi: 10.1590/s0100-879x2003001100005. Epub 2003 Oct 22.
Measles virus is a highly contagious agent which causes a major health problem in developing countries. The viral genomic RNA is single-stranded, nonsegmented and of negative polarity. Many live attenuated vaccines for measles virus have been developed using either the prototype Edmonston strain or other locally isolated measles strains. Despite the diverse geographic origins of the vaccine viruses and the different attenuation methods used, there was remarkable sequence similarity of H, F and N genes among all vaccine strains. CAM-70 is a Japanese measles attenuated vaccine strain widely used in Brazilian children and produced by Bio-Manguinhos since 1982. Previous studies have characterized this vaccine biologically and genomically. Nevertheless, only the F, H and N genes have been sequenced. In the present study we have sequenced the remaining P, M and L genes (approximately 1.6, 1.4 and 6.5 kb, respectively) to complete the genomic characterization of CAM-70 and to assess the extent of genetic relationship between CAM-70 and other current vaccines. These genes were amplified using long-range or standard RT-PCR techniques, and the cDNA was cloned and automatically sequenced using the dideoxy chain-termination method. The sequence analysis comparing previously sequenced genotype A strains with the CAM-70 Bio-Manguinhos strain showed a low divergence among them. However, the CAM-70 strains (CAM-70 Bio-Manguinhos and a recently sequenced CAM-70 submaster seed strain) were assigned to a specific group by phylogenetic analysis using the neighbor-joining method. Information about our product at the genomic level is important for monitoring vaccination campaigns and for future studies of measles virus attenuation.
麻疹病毒是一种高度传染性病原体,在发展中国家引发了重大的健康问题。该病毒的基因组RNA为单链、不分节段且呈负链极性。许多针对麻疹病毒的减毒活疫苗是使用原型埃德蒙斯顿毒株或其他当地分离的麻疹毒株研发的。尽管疫苗病毒的地理来源多样且采用了不同的减毒方法,但所有疫苗毒株的H、F和N基因在序列上具有显著相似性。CAM-70是一种日本麻疹减毒疫苗毒株,自1982年以来在巴西儿童中广泛使用,由Bio-Manguinhos生产。此前的研究已对该疫苗进行了生物学和基因组特征分析。然而,仅对F、H和N基因进行了测序。在本研究中,我们对其余的P、M和L基因(分别约为1.6 kb、1.4 kb和6.5 kb)进行了测序,以完成CAM-70的基因组特征分析,并评估CAM-70与其他现有疫苗之间的遗传关系程度。这些基因使用长距离或标准RT-PCR技术进行扩增,cDNA被克隆并使用双脱氧链终止法进行自动测序。将先前测序的A基因型毒株与CAM-70 Bio-Manguinhos毒株进行序列分析比较,结果显示它们之间的差异较小。然而,通过使用邻接法的系统发育分析,CAM-70毒株(CAM-70 Bio-Manguinhos和最近测序的CAM-70种子批次毒株)被归为一个特定的组。在基因组水平上了解我们的产品信息对于监测疫苗接种活动以及未来麻疹病毒减毒研究非常重要。