An optical biosensor with up-converting phosphor (UCP) marker is developed for the sensitive rapid immunoassay to the specific biomolecule. UCP can emit visible light when excited by infrared light. Through detecting and analyzing the content of UCP particles on the test strip after immunoreaction, the concentration of target analyte in the sample can be obtained. The detection sensitivity to plague IgG is better than 5 ng/ml; to plague FI-Ab is better than 100 pg/ml; to plague Yersinia pestis cell is better than 3*10^(4) CFU/ml. Good linear response characteristics and an excellent correlation (R2>=0.95) have been verified by quantitative detection results. In the practical application, detection results to 167 analytic samples have an excellent consistency with those obtained by reverse hemagglutination test. The up-converting phosphor technology (UPT) based biosensor has stable, reliable, and sensitive performances. It can meet the need of various bioassay applications.