Objective To investigate the detection capacity of esterase-3 (Es-3) in the laboratory animals monitoring laboratories in China, and to improve the quality management of laboratories. Methods We prepared the test samples according to the criteria of China National Accreditation Service for Conformity Assessment(CNAS), all the samples were certificated by homogeneity test and stability test. Then, samples with random numbers and standard operation instruction were distributed to the participant laboratories. The laboratories should submit their reports before the deadline expires. When the results are the same as the standard results, the laboratories will receive excellent remark; when the results are the same as the standard results except the hybridization type, the laboratories will receive satisfactory remark; otherwise, it will receive unsatisfactory remark. If a laboratory did not submit report, the laboratory will also receive unsatisfactory remark.Results Ten laboratories participated in the program, and no laboratory received excellent remark. Nine laboratories (90.0% of enrolled laboratories) had satisfactory results, while one laboratory (10.0% of enrolled laboratories) had unsatisfactory results. Conclusions The nationwide overall detection level of laboratories in Es-3 is relatively high. However, some details should be noticed and several laboratories should improve their detecting ability.