It has always been a challenging job to identify early pregnancy in buffaloes. Also, there may not be sufficient numbers of veterinarians to perform this task or cost of the service may be higher than that could be afforded. Buffaloes have a major contribution in milk production in Nepal. The productivity of the buffalo is poor mainly due to delay in first parturition and the subsequent conception time. BioPryn assay measures the blood level of Pregnancy Specific Protein B (PSPB), and this is considered as an early, accurate and easy to implement pregnancy diagnosis program in dairy cattle. However, its application in buffalo pregnancy diagnosis is yet to be established. The proposed study focused to find the basal and pregnant blood levels of PSPB in Nepalese buffaloes as a step towards validating this method in buffaloes. Based on this study, pregnancy based on the circulating PSPB level can be detected in buffaloes as early as week 3 post mating with BioPryn test. The research can be useful to veterinarians and farmers to make a vision in use of Biopryn test for early pregnancy diagnosis in buffaloes and to add information via research and actions in coming days.