Malarial infection is a common problem in tropical and subtropical countries. In current scenario P. falciparum malaria is a tough challenge to reduce infection in world wide. CD35 protein density on RBCs determines susceptibility to P. falciparum infection. An adhesion protein PfEMP-1 on surface of P. falciparum is responsible for adhering to CD35 a surface protein of RBCs. In our population, complement receptor Type-1 (CD35) gene polymorphism determine the density of CD35 protein on RBCs. As we know, CD35 protein is an adhesion protein for P. falciparum and high density confer chances to infection. For this study, we collected 270 blood sample (130, P. falciparum infected patient and 140, control) and estimate TNF-α level in both case and control population. Furthermore, we extracted genomic DNA and amplify the CR1 (CD35) gene of 1800 bp product, using thermal cycler. The PCR product was treated at SNP Intron 27 (T520C) with HindIII RFLP which generates 1300 bp and 500 bp. Polymorphism of CR1 (CD35) gene, SNP Intron 27 (T520C) in our population reveals three genotypes HH, HL and LL. Genotype HL (P=0.0365*) frequent distributed in population and showing association between case and control. H- Allele for High density and L-Allele for Low density. The frequency of H allele (in case=66.92%, control=33.07%) is high in compare to L alleles (in case = 58.74%, control = 41.25%). Frequency of H and L allele (P= 0.0484*) is also significantly associated with case and control population whereas, carriage rate were not associated (P=0.2287). Elevated TNF-α level in blood serum reveals quantity of infection level and its showing strong association (P<0.0001 ***) with P. falciparum malarial infection.