Early pregnancy diagnosis is an important management practice for reducing calving interval, increasing cattle reproductive efficiency, and the overall herd productivity. This study was undertaken to assess the viability of seed germination inhibition technique (Punyakoti test) for early pregnancy detection in cattle under the tropical rearing conditions. Twenty-four randomly selected cows were used for the experiment. Urine samples were collected and subjected to Punyakoti test, using maize seeds, within 6 hours of collection. Descriptive statistics employing means and standard error were used to analyse data, also, inferential statistics including analysis of variance and t-tests were employed to ascertain differences between the variables under study. Urine from pregnant cows had the highest (80.03±3.99) inhibitory effect while the distilled water group had the least (7.50±3.81) mean seed germination inhibition. The means of germinated maize seeds in pregnant and non-pregnant cow urine treatment groups were significantly different. The means of germinated maize seeds in unstripped and stripped pregnant cow urine were significantly different, while the mean shoot length values for the same treatments were not significantly different. Urine from 42 days’ pregnant cows had the highest germination inhibition (80.21±3.59) while the least (25.00±4.35) was observed on day 10. The Punyakoti test reliably detected pregnancy starting from 26 days after insemination. In conclusion, this technique can be used for detecting pregnancy as early 26 days after insemination. The potential role of steroids and involvement of abscisic acid (ABA) in seed germination inhibition requires further investigation.