Combined detection and genotyping of Chikungunya virus by a specific reverse transcription-polymerase chain reaction
Combined detection and genotyping of Chikungunya virus by a specific reverse transcription-polymerase chain reaction
复制标题
DOI：
10.1002/jmv.10085
复制
发表时间：
2002-07-01
影响因子：
12.7
通讯作者：
Igarashi, A
中科院分区：
文献类型：
--
作者：
Hasebe, F;Parquet, MC;Igarashi, A
建立了一种检测基孔肯雅病毒感染的逆转录聚合酶链反应（RT-PCR）方法。以基孔肯雅病毒非结构蛋白1 （nsP1）和糖蛋白El （El）基因为基础，设计了两组引物。从1953年在坦桑尼亚分离的S27原型病毒接种白纹伊蚊C6/36细胞和1998年在马来西亚分离的MALh0198、MALh0298和MALh0398株细胞培养液中提取总RNA。对于这两组RNA样本，分别从nsP1和El基因中有效扩增出预期的354和294碱基对（bp） cDNA片段。利用部分El基因序列数据，对马来西亚毒株和从世界基孔肯雅流行不同地区分离的其他病毒毒株进行了系统发育分析。在1998年流行期间分离出的马来西亚菌株与亚洲基因型的其他成员归为一类。(C) 2002 Wiley-Liss, Inc。
A reverse transcription-polymerase chain reaction (RT-PCR) was developed for the detection of Chikungunya virus infection. Based on the nonstructural protein 1 (nsP1) and glycoprotein El (El) genes of Chikungunya, two primer sets were designed. Total RNA were extracted from the cell culture fluid of Aedes albopictus C6/36 cells inoculated with the S27 prototype virus, isolated in Tanzania in 1953, and the Malaysian strains (MALh0198, MALh0298, and MALh0398), isolated in Malaysia in 1998. For both sets of RNA samples, the expected 354- and 294-base pair (bp) cDNA fragments were amplified effectively from the nsP1 and El genes, respectively. Phylogenetic analysis was conducted for the Malaysian strain and other virus strains isolated from different regions in the world endemic for Chikungunya, using partial El gene sequence data. The Malaysian strains isolated during the epidemics of 1998 fell into a cluster with other members of the Asian genotype. (C) 2002 Wiley-Liss, Inc.