Temperature detection is particularly important in the medical and scientific fields. Although there are various temperature detection methods, most of them focus on broad temperature detection, and basic research in specific fields, especially the detection of subtle temperature changes (32–34 °C) during wound infection, is still insufficient. For this purpose, a novel colorimetric temperature sensing probe is designed in this paper, which can quickly and intuitively respond to small temperature changes within a specific range through color changes. In this paper, hyperbranched polyethyleneimine (HPEI) was modified by isobutyrylation to prepare hyperbranched temperature-sensitive polymer (HPEI-IBAm). And it was combined with gold nanoparticles (AuNPs) prepared by a sodium citrate reduction method to construct an HPEI-IBAm-AuNP colorimetric probe. The probe exhibits excellent stability, even at salt concentrations of up to 12 g/L, thanks to the abundant amino functional groups and the large steric hindrance effect unique to HPEI-IBAm. In particular, the temperature detection range of the probe is precisely locked within 32–34 °C, enabling it to respond quickly and accurately to small temperature changes of only 2 °C. This feature is perfectly suited to the practical needs of temperature detection in infected wounds. The linear fitting coefficient of the temperature response is as high as 0.9929, ensuring the accuracy of the test results. The detection performance of the probe remained highly consistent over 10 cycles, fully proving its excellent reusability and durability. In addition, a flexible colorimetric sensor was prepared by combining the probe with polydimethylsiloxane (PDMS) film. This sensor is capable of rapidly detecting human skin temperature in real time, achieving an accuracy of 99.07% to 100.61%. It can provide a possible solution to the challenges of delayed and difficult temperature detection caused by different body parts and uneven surfaces, among others. This demonstrates its extensive practical value and potential, and it is expected to be further applied in the monitoring of wound infections.