In order to explore the genotype distribution of human enterovirus group B (HEV-B) in Shandong Province and to study the correlation between HEV-B serotypes and disease outbreaks, we sequenced and analyzed the entire VP1 coding region of HEV-B isolated from acute flaccid parolysis (AFP) system and disease outbreaks in Shandong province during 1998-2008. All together twenty nine HEV-B serotypes were identified, including twenty Echovirus (ECHO) serotypes, five Coxsackievirus B (CVB1-5) serotypes, one Coxsackievirus A9(CVA9) serotype, and newer enteroviruses EV73, EV75, and EV97. E11, CVB3, E6, E14 and E25 were the five frequently isolated serotypes from AFP surveillance system. CVB3, CVB5 and ECHO30 were the major causative agent of aseptic meningitis in Shandong province. Comparison of nucleotide sequence homology showed 75.4%-99.6% inter-typic identities within Shandong strains, and 73.8%-85.2% identities with prototype strains. Amino acid sequence comparison showed the differentiation was not much. Our research showed different serotypes possessed distinct time-cycling pattern, and different sub-genotypes could be further classified according to the inter-typic genetic distance. Thereby the route and range of transmission of HEV could be determined.