乙脑病毒基因Ⅰ、Ⅲ型特异性全基因组引物

The genotype-specific primers for amplying and sequencing the genotype Ⅰ ＆ Ⅲ Japanese encephalitis virus

目的为构建高通量乙脑病毒全基因组序列测定平台，设计两套分别针对基因Ⅰ型和基因Ⅲ型乙脑病毒全基因组的型特异性引物。方法根据GenBank公布的乙脑病毒全基因组序列为参考信息，设计了两套基因Ⅰ型和基因Ⅲ型乙脑病毒的基因型特异性扩增测序引物。并由其测定本研究室采集并分离得到的121株乙脑病毒全基因组序列，含63株基因Ⅲ型乙脑病毒，58株基因Ⅰ型乙脑病毒。结果设计得到基因Ⅲ型乙脑病毒全基因组扩增测序引物17对，基因Ⅰ型乙脑病毒型特异性扩增测序引物16对。使用两套引物完成121株乙脑病毒全基因组序列的测定。测定完成的基因Ⅰ型乙脑病毒全基因组序列平均值为40．037，基因Ⅲ型乙脑病毒全基因组序列平均质量值为40．857。结论本研究设计的两套型特异性引物能高效特异地完成基因Ⅰ型和基因Ⅲ型乙脑病毒全基因组序列的扩增，为构建高效稳定的乙脑病毒全基因组序列测定平台奠定了基础。

Objective For constructing the high-throughput platform of sequencing the JEV whole genome, the two systems of multiplex primers for genotype Ⅰ and Ⅲ should be designed and used for detected the whole genome of genotype Ⅰ and Ⅲ JEV in the research. Methods The two systems of JEV genotype-specific primers was designed based on the reference sequence of all the available genotype Ⅰ and Ⅲ JEV genome sequence on GenBank, then, they were used to amply and sequence the 121 JEV strains isolated in China contains 63 Gill JEV and 58 GI JEV. Results The self-designed genotype-specific primers for genotype I and genotype m JEV were 16 pairs and 17 pairs respectively, which were used for detecting the whole genome of 121 JEV. The average quality value for GI JEV is 40.037. The average quality value for Gm JEV is 40.857. Conclusion The two systems of JEV genotype-speeific primers could sequenced the genotype Ⅰ and Ⅲ JEV qualified and specific. It is the basis of the high throughput platform of sequencing the JEV whole genome.