A novel carbon dot/Rhodamine B-based ratiometric fluorescent probe was developed for a highly sensitivity and selective detection of nitrite (NO2-). The probe showed colour changes from blue to orange under ultraviolet light in response to NO2- with a detection limit as low as 67 nM in the range of 0 to 40 μM. A ratiometric fluorescent test paper was successfully prepared using the probe solution, which demonstrated its feasibility towards a rapid and semi-quantitative detection of NO2- in real samples.