Objectives:This study aimed to assess the knowledge of junior doctors on the 2015 chronic hepatitis B (CHB) guideline recommended by Chinese society of infectious diseases and Chinese society of hepatology, Chinese medical association. Methods: Junior doctors, who were already engaged in the field of CHB, were invited to complete a questionnaire-based survey between May and June 2016. The questionnaire consisted of 28 items focusing on knowledge in the following three sections: motherto-child transmission (MTCT) prevention, response-guided therapy (RGT) strategies, and special patients' antiviral therapy. Results: Responses were received from 562 out of 600 participants. Among those 562 junior doctors, about 10%~30% were not familiar with the use of hepatitis B vaccine and hepatitis B immunoglobulin (HBIG), and 18.8% did not know that newborn with HBsAg-positive mother could be breastfed after giving HBIG and vaccine. About 20.6% of junior doctors did not know the possible withdrawal indications recommended by the guideline. For HBeAg-positive patients receiving low genetic barrier drugs, about 30% of junior doctors did not know that the antiviral strategies should be adjusted according to HBV DNA levels at treatment week 24. For CHB patients receiving chemotherapy, about 25% of participants did not know antiviral therapy should be started at least 1 week earlier, and about 20% did not know that adefovir dipivoxil or tenofovir disoproxil fumarate should be avoided in patients with kidney diseases or high-risk of developing kidney diseases. Conclusions: The knowledge on CHB guideline was rather unsatisfactory in junior doctors. Our finding highlights the urgent need for strengthening junior doctors to gain a greater understanding of CHB guideline.