Aedes albopictus is an important vector of many mosquito-borne viral diseases, including dengue fever and Zika. In recent years, it has spread and colonized tropical, subtropical and temperate regions worldwide. Monitoring of Ae. albopictus population dynamics is an important tool for early warning of mosquito-borne infections. Because the genetic diversity and genetic structure of natural populations are the genetic bases of population dynamics, studies of population genetics can reveal the origin, differentiation and dispersal characteristics of Ae. albopictus populations. Then, their evolutionary potential and environmental adaptability can be analyzed, providing a theoretical basis for the formulation of accurate Ae. albopictus surveillance and integrated control programs. In 2018, 552 Ae. albopictus larvae were collected during an invasive mosquito species surveillance project in China's Yangtze River Basin. Morphological analysis was performed to assign the adult mosquitoes to species, and then the genetic marker ITS2 was amplified and sequenced. There were 179 haplotypes among 552 ITS2 sequences. In total, 155/179 (86.59%) haplotypes were specific to individual populations, and 24/179 (13.41%) haplotypes were shared by populations. Hap4 (126), Hap7 (43), and Hap16 (34) were the most numerous haplotypes and the most widely distributed. The overall Hd was 0.928, π was 0.031, the mean nucleotide difference number (K) was 7.255, and the number of segregating sites was 169. TCS network maps mainly showed a single star-like scattered distribution. According to geographical location, there were no obvious haplotype groups, and the haplotypes were intricately connected. The genetic diversity of Ae. albopictus populations in the Yangtze River Basin was high. The molecular variance observed in the populations of Ae. albopictus mainly occurred among individuals within populations, accounting for 98.79% of the total, while that among populations accounted for only 1.21% of the total. Only the populations of Ae. albopictus in the Chongqing and Sichuan regions showed a moderate degree of population genetic differentiation, while genetic differentiation between the other regions were small, gene exchange was very common, and genetic differentiation within populations was minimal. According to this study, the genetic diversity of Ae. albopictus populations in the Yangtze River Basin is high, the genetic differentiation among populations is small, and gene exchange is common. In addition, frequent interregional exchange exacerbates the abnormal spread of vectors. This study highlighted the potential spread route of the vector Ae. albopictus in the Yangtze River Basin. There are three potential dispersal routes for Ae. albopictus populations in the Yangtze River Basin. The findings could be helpful for effective surveillance and early warning of Ae. albopictus vectors.