Hepatitis B virus (HBV) is a small, circular, partial double stranded DNA virus classified under hepadnaviridae. Almost more than 350 million HBV infected persons were reported universally. Blood/its product transfusion represents one of the commonly routes of HBV transmission that may increase the HBV infection globally. HBV is diagnosed routinely by detection of both HBV specific antigens (HBsAg, HBcAg, HBeAg) and antibody markers (HBcAb). ELISA and Electrochemiluminescent immunoassay (ECLIA) are the most common methods to detect HBV antigens or antibodies. This study aimed to compare between ECLIA (Cobas e 411) and ELISA for detecting HBsAg and Total HBcAb among blood donors at National Blood Transfusion Center-Taiz branch (NBTRC-TB). 125 blood samples that were tested by ECLIA were chosen randomly and re-tested using ELISA method. The result of HBsAg seropositivity by ELISA method appeared as 28/125 (22.4%) compared to 25/125 (20%) by ECLIA method; whereas the result of total HBcAb seropositivity by ELISA method appeared as102/125(81.6%) compared to 100/125(80%) by ECLIA method. The current study appears to be a rapprochement between the results of two methods (ELISA and ECLIA) for detection HBsAg and HBcAb). In abscess of ECLIA, ELISA could be a good technique for blood screening before transfusion. Using molecular methods such as PCR will be more a curate to ensure blood safety before transfusion.