Norwalk-like viruses(NLVs) antigen detection ELISA was newly developed. It is constructed with monoclonal antibodies(MAbs) as a capture antibodies and hyperimmune rabbit sera as a detector antibodies. The two MAbs reacted broadly and specifically with genogroup I and genogroup II NLVs, respectively. Total detection time is less than 3 hours and at least 90 stool samples could be tested at one time. The concordance rate with ELISA and RT-PCR was 69% and it could be raised to 71% if capsid primer sets were used. Epidemiological study revealed that among 664 normal persons one sample(0.2%) was confirmed positive by both ELISA and RT-PCR, suggested that NLV carrier may exist in the normal person. The ELISA kit has advantages such as rapid, simple and multi samples detection, and useful for the first screening test. Also it is big tool for epidemiological surveillance of NLVs invasions.