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Paratuberculosis (PTB) is a chronic disease affecting ruminants caused by mycobacterium avium subspecies paratuberculosis (MAP), characterized by the iceberg phenomena, as many cases are subclinical and underdiagnosed. An early diagnosis is imperative; however, no reliable single test is available, leading to delayed culling. Furthermore, using more than one test increases the rate of positively diagnosed cases. The current study aimed to detect subclinical PTB in dairy cows in El-Minia governorate, Egypt, using ELISA jointly with the detection of MAP by PCR. The positive cases were also subjected to pathological examination to determine whether lesions were present and their severity. A total of 145 cows of different breeds (Baladi, Mixed, and Holestins) and ages were tested by ELISA and PCR. Our results showed that the positive PTB cases detected by ELISA and PCR were 17.24% and 20%, respectively. Holestins have a significantly higher infection rate 31.70% than Baladi and Mixed breeds. In contrast, the native (Baladi) breed has the lowest infection rate 11.47%. Moreover, PTB is more common in cows aging (age ≥ 1.5-