Abstract：Sequence alignment analysis was conducted on domestic Tomato yellow leaf curl virus （TYLCV）by DNA-A genomic sequence．The result of nucleotide sequence blast showed that the complete nucleotide sequence of various China isolates shared high identity from each other．Phylogenetic relationship analysis of TYLCV full genomic and coat protein nucleotide and amino acid sequence reveals that the middle eastern and coastal tomato production regions of China were infected by TYLCV-IL isolates．The Western inland areas and Southwestern inland areas were infected by TYLCCNV isolates．Gene polymorphism analysis showed that TYLCV had a strong conservative evolution，and the distribution of nucleotide mutations（Pi）on the chromosome with high polymorphism．Therefore，according to strict conservative region of TYLCV，3 pairs of specific primers were designed，and a high sensitive，high efficient and with high specificity multiplex PCR detection method of TYLCV was etablished.
