PCR-RFLP技术用于中国登革2型病毒株基因型快速分型

PCR-RFLP for typing of Chinese DEN-2 genotypes

利用RT PCR的方法 ,对中国 5株登革 2型病毒的prM E基因进行扩增 ,选择合适的限制性内切酶将扩增产物特异地切割成若干长度不同的片段 ,经 5 %聚丙烯酰胺凝胶电泳后 ,对获得的银染带型进行分析。结果表明FJ 10株和FJ 11株属同一基因型 ,D2 4 3株和D2 4 4株属同一基因型 ,D2 0 4株与上述 4株基因型不同 ,这与核苷酸测序分析结果完全一致。在此基础上建立PCR RFLP技术用于登革 2型病毒基因型快速分型 ,具有省时、操作简单、不需使用同位素等特点 。

In this study, the prM-E genes of 5 Chinese DEN-2 isolates from Fujian, Guangxi and Hainan were amplified by RT-PCR.Two restriction enzymes were selected by software DNASTAR to generate the polymorphisms in different restriction fragments length for strain discrimination. The digested fragments were visualized by silver staining. The results showed that strain DEN-2 FJ-10 and strain FJ-11 belong to the same genotype,strain D2-43 and strain D2-44 are another genotype and strain D2-04 is different from the former 4 strains. Those results by PCR-RFLP correlate well to those previously determined by sequencing. The PCR-RFLP technology is a useful tool for rapid typing of viral isolates, especially suitable for laboratory test and clinical diagnosis in endemic areas of dengue fever.