Osteoporosis is a bone disease, characterized with bone tissue deterioration and decreasing level of bone mineral. Appropriate diagnostic is mandatory to prevent osteoporosis and provide a space for enough surgical and pharmacologic treatments. Available radiological techniques cause various health issues and research proved that biological markers are playing a major role for bone mineralization, can be a good alternative for diagnosing osteoporosis. C-terminal telopeptide (CTx) has been found as a potential biomarker, this study uses interdigitated electrode (IDE) sensor to identify CTx. Carbon nanofiber (CNF) measured as 50 nm by high-resolution microscope was used to modify the sensing electrode surface through amine-linker and it promotes higher anti-CTx antibody immobilization. CTx was quantified on this antibody anchored surface and attained the limit of detection to be 25 pg/mL. A regression co-efficient [y = 0.566x − 0.2627, R2 = 0.9823] with triplicates (n = 3) determined the high-performance of IDE. Control molecules failed to increase the current flow, is indicating the selective CTx detection and CTx serum spiked human serum provided an increment in current–volt at linear concentrations are from 25 to 800 pg/mL, indicating the selective detection.