A probe system for real-time quantitative and qualitative analysis of a biomaterial, and a reaction chamber with the probe system, and an analysis method thereof are provided. The probe system, which is included in the reaction chamber having an optically transmissive flat bottom surface and having a test sample accommodated therein, includes a target probe-reporter probe linker accommodated in the reaction chamber and including a target probe, which includes a sequence complementary to a target nucleic acid sequence to be detected, a first fluorophore and a first quencher, and a reporter probe linked to an end of the target probe and including a sequence non-complementary to the target nucleic acid sequence, and a capture probe included in a biochip formed on a bottom surface of the reaction chamber and including a complementary sequence hybridizable with the non-complementary sequence of the reporter probe, a second fluorophore and a second quencher.