对大型黄病毒PCR产物进行直接测序，以分析基因组变异和开展分子流行病学调查。
Direct sequencing of large flavivirus PCR products for analysis of genome variation and molecular epidemiological investigations.
作者信息
Lewis J G, Chang G J, Lanciotti R S, Trent D W
机构信息
Division of Vector-Borne Infectious Diseases, Centers for Disease Control, Fort Collins, CO 80522.
出版信息
J Virol Methods. 1992 Jul;38(1):11-23. doi: 10.1016/0166-0934(92)90165-a.
The polymerase chain reaction (PCR) was used to amplify viral cDNAs from selected regions of dengue genomic RNA by using appropriate 'consensus' primers. DNA amplicons containing the structural genes from all 4 dengue serotypes were prepared and directly sequenced using dengue-virus-specific primers. This method can characterize reliably flavivirus field isolates at the molecular level without extensive virus propagation and molecular cloning, and will be a valuable tool for molecular epidemiological studies.
采用聚合酶链反应（PCR），通过使用合适的“共有”引物，从登革热基因组RNA的选定区域扩增病毒cDNA。制备包含所有4种登革热血清型结构基因的DNA扩增子，并使用登革热病毒特异性引物直接测序。该方法无需大量病毒繁殖和分子克隆，就能在分子水平可靠地表征黄病毒野外分离株，将成为分子流行病学研究的宝贵工具。
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