A reflective fiber-optic Fabry-Perot cavity probe sensor is proposed to selectively measure cholesterol concentration by insert single mode fiber into ceramic tube and immobilize epoxy resin (ER)/graphene oxide (GO)/ beta-cyclodextrin (beta-CD) multi-layer film onto end face of ceramic tube. EDC/NHS activated GO is selected to form chemical binding with beta-CD, and beta-CD is the sensitive materials to bind with cholesterol molecules. With multi-layer film assisted, the sensitivity of sensor to cholesterol concentration can reach 3.92 nm/mM and the limit of detection reaches 3.48 mu M. In addition, 4 mM hemoglobin, glucose and ascorbic acid are doped into a set cholesterol sample and verified the highly selectivity of sensing cholesterol. Furthermore, the reproducibility was proved by measure the spectrum of four sensors with same fabrication process, and the reusability was also proved by repeated measurements. Overall, the sensor features with high mechanical strength, ease of fabrication, real-time monitoring, low cost and ease for measurement that given by probe structure. Therefore, the sensor provides a remarkable analytical platform for biosensing applications.