This Study aimed to comparison between diagnostic methods to evaluated the prevalence of Vibrio cholerae .A total of 90 samples include  40 birds faeces  were collected from bird farms and birds breeding stores for Ducks, chickens, Pigeons, Pet birds  and  50 hand swab from owners of birds (Males and Females) .collection from different places Al-hurya city, Abu Ghraib city, AL-Yarmouk area ,University district area in  Baghdad  Province, at the period of February 2021 to February 2022, in order to evaluate the routine laboratory diagnostic procedures in the diagnosis of V. cholerae isolates and compared them with molecular methods as Polymerase chain reaction (PCR).V.cholerae has been isolated and identified by using culturing method in addition to biochemical tests, Vitek2 system. The results showed that in culturing method percentage of isolation (43.33%)from birds feces, and (33.33%) in hand swabs,in  biochemical tests(33.33%) in birds feces, and (10%)from  hand swabs in Vitek2 system  (6.7%)from each birds feces and  hand swabs. PCR technique was used to detect sodB Gene encoding to outer membrane protein of V. cholerae ,on the PCR results, the rate of V. cholerae isolation from birds feces samples was 23.3 %. and 6.66% from  hand swab samples,in Comparison  PCR results showed that there were high sensitivity 100% with cultural, biochemical test , while 44% with Vitek2 system respectively.in presnt study found that  diagnosis of V.cholerae by routine diagnostic may be misidentifications and using of PCR more faster, accurate ,sensitive and  specific for correct identification.