Babesia sp are the intra-erythrocytic protozoan parasite causes Bovine babesiosis. Babesia bovis and Babesia bigemina are two species that commonly infest cattle. A conventional parasitological technique is commonly used to diagnose Babesia sp. This technique, however, has limitations in subclinical infections. Polymerase chain reaction (PCR) has a high sensitivity and can identify parasites at much lower concentrations. The goal of this study was to detect B. bovis and B. bigemina in blood cattle and buffaloes from fields using the conventional Giemza Staining Thin Blood Smear method (GSTBS). In addition, the study compared the effectiveness of a single and duplex PCR in detecting parasites. One hundred and ninety-eight blood samples from cattle and buffalo were collected in Bogor (West Java), Pemalang, and Brebes (Central Java). The parasite's existence was determined using a thin blood smear. Furthermore, the PCR assay was employed with primers Bovar 2A and Bg3/4 specific for B. bovis and B. bigemina, yielding amplified products of 166 and 689 bp, respectively. Based on parasitological findings, 14 of 198 blood samples (7.1%) tested positive for Babesia sp. The results of a single PCR demonstrated that 21,21% (42/198) of the samples were positive for B. bovis, whereas duplex PCR successfully identified B. bovis and B. bigemina in 72 samples (36,36%). This finding indicated that duplex PCR is more efficient and cost-effective than GSTBS and single PCR for surveying babesiosis in epidemiological studies.