Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease caused by the SFTS virus (SFTSV). Hubei Province is a major epidemic area for SFTS in China. In this study, quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and serological testing (IgM) were used simultaneously for laboratory detection of SFTS; however, testing results showed poor consistency between these two methods. Further analysis revealed that time post-onset was the main factor leading to inconsistent results. Thus, qRT-PCR is unable to detect all SFTS cases, and serological testing is essential. Here, 15 strains of SFTSV were successfully isolated from serum samples of acute SFTSV infection and their complete genomes were sequenced and submitted to GenBank. Phylogenetic analysis showed that the 15 SFTS virus strains clustered into four independent genotypes (A, B, D, and E), demonstrating that at least four genotypes of SFTSV have been co-circulating in Hubei Province. Furthermore, four strains of our isolates (HB2014-31, HB2014-35, HB2014-36, and HB2014-37) clustered in genotype E, which was the predominant genotype in Japan and South Korea. In this study, we identified multiple co-prevalent genotypes and confirmed the existence of genotype E viruses circulating in the Dabie Mountains of Hubei, central China. We concluded that SFTSV strains from Hubei exhibit most of the genetic diversity found in China.