Early pregnancy diagnosis is an essential tool in successful reproductive management of ruminants. We attempted to explore the effectiveness of a marketable ELISA test kit for assessment of Pregnancy Associated Glycoproteins (PAGs) in peripheral blood for early pregnancy identification in cattle, buffalo, goats and sheep. A total of 120 blood were taken from jugular vein of different breeds of cattle (Achai, Achai x Jersey, Holstein Friesian and Jersey), buffalo (Nilli Ravi, Aza Kheli and non-descript), goats (Beetal, Teddy and non-descript) and sheep (Bulkhi, Karri and non-descript). In cattle, the average sensitivity, specificity, false pregnancy prognostic value, false non-pregnancy prognostic value and accuracy of the PAG-ELISA test were 86.77%, 66.67%, 86.77%, 100% and 90%, respectively. Similarly, the current study indicated that in buffalo, average sensitivity, specificity, false pregnancy prognostic value, false non-pregnancy prognostic value and accuracy of the PAG-ELISA test were 62.22%, 74.6%, 62.22%, 88.57% and 76.67% respectively. The current study demonstrated that average sensitivity, specificity, false pregnancy prognostic value, false non-pregnancy prognostic value and accuracy of the PAG-ELISA test were 100%, 100%, 100%, 100% and 100% respectively in goat. Similarly, the results in sheep showed that the average sensitivity, specificity, false pregnancy prognostic value, false non-pregnancy prognostic value and accuracy were 95.23%, 91.67%, 95.23%, 100% and 100% respectively. The results showed that PAG is a valuable marker for early pregnancy diagnosis in ruminants.