Background: We have recently shown that the copy number of salivary amylase (AMY1) gene was significantly decreased, and the obesity-related salivary biomarkers resistin, MCP-1, TNF-α, IL-6, and CRP were significantly increased in overweight/obese children compared to normal weight. This study aimed to evaluate the association of AMY1 copy number variant (CNV) with obesity and inflammatory markers. Seventy-six participants aged between 6 and 10 years have participated, and the saliva samples were collected along with the anthropometric measurements. Methods: AMY1 copy number was analyzed by 3D digital PCR, and obesity-related biomarkers were performed with a Bioplex multiplex analyzer. Results: The mean AMY1 copy number was higher in normal weight (7.90 ± 0.38) compared to the overweight/obese group (6.20 ± 0.29). The association of AMY1 CNV with obesity and inflammatory markers showed significant negative correlation [CRP, β = −0.238 (p < 0.05); resistin, β = −0.25 (p < 0.05); MCP-1, β = −0.304 (p < 0.01)] except for complement factor D, TNF α and IL-6. The anti-inflammatory cytokine, IL-10 reported a positive correlation with AMY1 copy number with a β = 0.268 (p < 0.05). The multivariable model adjusted with age and gender depicted a similar correlation with obesity markers. Conclusion: Our results report that AMY1 CNV is associated with obesity and inflammatory biomarkers in children's saliva sample.