The quantitative buffy coat (QBC) technique and conventional Giemsa thin blood smear was compared to determine the sensitivity and specificity of the technique in detecting blood parasitic infection of the rodent populations from four urban cities in Peninsular Malaysia. A total of 432 blood samples from four rat species (Rattus norvegicus, Rattus rattus diardii, Rattus exulans and Rattus argentiventer) were screened using both techniques and successfully detected two blood protozoan species (Trypanosoma lewisi and Plasmodium sp.) with Trypanosoma lewisi predominantly infecting the population. Results showed that Giemsa-stained thin film (GTF) was the better detection method on blood parasitemia (46.7%) compared to Quantitative Buffy Coat method (38.9%) with overall detection technique sensitivity and specificity at 83.2% and 74.8% respectively. The sensitivity in detection of Trypanosoma lewisi was 84.4% with value slightly lower for Plasmodium sp. infections at 76.6%. Statistical analysis proved that GTF technique was significantly more sensitive in the detection of blood protozoan infections in the rodent population compared to QBC (p<0.05).