Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of Johne's disease. The ability to detect MAP rapidly and accurately is important for cattle breeding. The present study compares the results under real time PCR technique on individual feces and on pools of 2 feces on the one hand to the results under coproculture considered as the reference method on the other hand.One hundred bovine fecal samples were selected by the laboratory (LDAR87) for MAP detection. Individual fecal samples were analyzed under real time PCR and were used to create pooled fecal samples. The 100 samples were randomly pooled by two and analyzed by PCR. 31 fecal samples, which were positive under coproculture, were mixed with negative coproculture feces. For each experiment, 1g of feces was used. DNA was extracted from fecal suspension using the ADIAPURE® kit (Adiagene). PCR was performed using the ADIAVET PARATB PCR® kit (Adiagène).While 31% of the samples were positive under coproculture, 34% of the samples turned out to be positive under PCR on individual samples and 36% of the samples under PCR on pools of 2 samples.The PCR test results concurred with the coproculture results. Under PCR tests on pooled samples more feces where positive than under PCR on individual samples. The differing results concerned weakly contaminated feces. These differing results could be due to the heterogeneity of the feces.These data indicate that the use of real time PCR on pooled fecal samples is as effective as coproculture to detect MAP without decrease of sensitivity.