Because of the high prevalence of Hepatitis B virus (HBV), HBsAg tests play an extremely significant role in donor screening in China. In our multicentre study, the HBsAg assays used in blood screening were evaluated for their analytical sensitivity and the ability to detect mutants. A panel of eight samples was sent to 85 blood banks. The panel included four HBsAg-positive serum samples at concentrations of 0·16, 0·46, 0·9 and 1·73 IU mL−1; three recombinant HBsAg mutants (G145R, T131I and K141E) with defined concentrations and one negative sample. All laboratories were required to detect the samples with their routine procedures.All the 85 blood banks reported their results before the closing date, and 170 data sets were submitted. The wild-type samples at concentrations of 1·73 and 0·9 IU mL−1 were correctly identified as positive in all data sets, whereas the other two samples (at concentrations 0·16 and 0·46 IU mL−1) were not consistently detected as positive. Regarding the mutants, significantly different results were reported among all the assays involved. All users of Hepanostika HBsAg Ultra and most users (16/17) of Abbott Murex HBsAg Version 3 detected all mutant samples, whereas some of the commercial assays failed to detect any of the three mutant samples. Some HBsAg assays with unsatisfactory sensitivities and capabilities of mutant detection were used by a large percentage of blood banks, which might lead to a high transfusion risk in China. A list of assays of high sensitivity and good ability of mutant detection should be recommended for use in blood screening.