Detection of Helicobacter pylori is essential for the prevention of gastric cancer. By detecting the metabolized NH 3 , it was able to noninvasively reveal the state of H. pylori; however, it is still a challenge since the metabolized NH 3 concentration is much lower for conventional gas sensors. Herein, we developed a MoS 2 -based NH 3 sensor for continuous, real-time monitoring of H. pylori growth. The atomic thin layer and the all-exposed surface of MoS 2 facilitate NH 3 adsorption and charge transferring. A high-response NH 3 sensor was prepared by surface decoration of MoS 2 by depositing metal nanoparticles. The Fe-decorated MoS 2 sensor outperformed with a high response of 40.9% for 5.7 ppm of NH 3 at 25 ± 2 °C, low LOD (6.2 ppb), and long-term stability with a response of 12.5% for 5.7 ppm of NH 3 after 5 months. The Fe-decorated MoS 2 sensor was applied to the detection of H. pylori and the real-time in situ monitoring of its 92 h growth cycle. The NH 3 release curve of the exponential phase during H. pylori growth was continuously monitored, and the NH 3 concentration was quantified. The maximum specific rate of NH 3 release was 0.195 ± 0.005 h −1 , which is well-consistent with the nature of H. pylori growth. This study opens up a technological roadmap for noninvasive detection of H. pylori in the future.