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A study from Thailand showed no significant association between the adiponectin (ADIPOQ) gene rs1501299 polymorphism and knee osteoarthritis (OA) risk. To investigate this association in a Chinese population, we conducted this case–control study involving 372 knee OA patients and 453 controls. Genotyping via standard PCR and restriction fragment length polymorphism (PCR-RFLP) showed that TT genotype (TT vs. GG: adjusted odds ratio (OR) (95% confidence interval (CI)) = 1.70 (1.01–2.86)) or T allele (T vs. G: adjusted OR (95% CI) = 1.26 (1.02–1.56)) of ADIPOQ gene rs1501299 polymorphism significantly increased the risk of knee OA. Significant associations were also observed in subgroups ≥55 years (TT vs. GG: adjusted OR (95% CI) = 2.21 (1.00–4.86)) and body mass index (BMI) < 25 kg/m2 (TT+GT vs. GG: adjusted OR (95% CI) = 1.53 (1.03–2.29)), but not in the subgroup analysis of sex. In conclusion, the ADIPOQ gene rs1501299 polymorphism intensifies the risk of knee OA in this Chinese Han population. Nevertheless, further studies with larger sample sizes in other populations are warranted to verify this finding.