Genetic diversity and relationship with litter size were analyzed in Hu sheep using 6 microsatellite markers LSCV043, BMS2508, GC101, 300U, Bulge5, and 471U, which were closely linked to FecB gene. Thirty-four alleles and 53 genotypes were detected at 6 microsatellite loci, which were all polymorphic. Three markers LSCV043, BMS2508 and 300 U were highly polymorphic loci. The polymorphism information content (PIC) were 0.6674, 0.6035, and 0.5615, respectively. The total mean of litter size of genotype LSCV043 107 bp/123 bp was significantly higher than that of genotype 110 bp/123 bp. The litter size of the first lambing for genotypes BMS2508 154 bp/154 bp, 154 bp/170 bp and 154 bp/200 bp were significantly higher than that of genotype 170 bp/170 bp (P<0.05). Furthermore, total litter size for BMS2508 170 bp/170 bp was the lowest in all the genotypes within this locus (P<0.05). No significant difference was detected in other genotypes.