One of the major sources of economic loss in dairy herds is Mastitis. The present study aimed at analyzing SNPs in the CXCR2 gene in river buffalo (Egyptian breed) and their association with mastitis. DNA from blood was extracted from thirty buffaloes, 15 healthy and 15 with mastitis and CXCR2 gene (1923bp) was amplified. The nucleotide analyses revealed the presence of 17 single nucleotide polymorphisms (SNPs) (11 mutual and 6 non-mutual) from which 16 SNPs were novel. Three mutual SNPs were associated with mastitis buffalo with genotypes CT and TT at both c.1-g.139(C>T) and c.546(C>T); GG at c.3+g.229 (A>G) were susceptible to mastitis. Buffalo with CC at both c.1-g.139 and c.546; AG and AA at c.3+g.229 were resistant to mastitis. On the other hand, six non-mutual SNPs were associated with mastitis. Healthy buffaloes were TT and CC homozygous at SNP c.1-g.53 (T>A) and c.127 (C>A), respectively. Mastitis buffalo were CC, TT, GG, CC homozygous at SNPs c.1-g.103 (A>C), c.779(C>T), c.784 (A>G), c.790 (A>C), respectively. Overall, the results showed that CXCR2 can be a valuable candidate gene for the prediction of disease resistance in buffalo.