Objective To explore the application value of a novel portable endoscope to perform upper gastrointestinal tract examinations in primary medical units. Methods A total of 532 subjects receiving portable endoscope examination were enrolled for analysis. The primary outcome was the success rate of operation. The secondary outcomes were the operation time, examination results, polyp removal and biopsy pathology results, and the subjective evaluation. Results In 532 cases, 2 were withdrawn midway after the endoscope was inserted into the esophagus due to the patients’ inability to tolerate the examination. Additionally, 6 cases did not undergo examination of the descending part of the duodenum because of serious reactions during the procedure. Ultimately, 524 cases successfully completed the upper gastrointestinal examination, and the success rate was 98.5%. The average examination time was (4.7±1.8) min, and the average time for disposal sheath wearing and removing was (4.2±1.4) min. The most common lesions were chronic non-atrophic gastritis (85.1%, 451/530), reflux esophagitis (14.7%, 78/530) and bile reflux (14.0%, 74/530). A total of 10 cases of polyp removal were completed, and the polyp removal rate was 71.4% (10/14). Biopsy pathological diagnosis was completed in 44 cases, and the biopsy rate was 8.3% (44/530). The main discomfort symptoms during the examination were nausea (53.6%, 285/532), vomiting (51.1%, 272/532), and sore throat (38.5%, 205/532), the main discomfort symptoms after the examination were sore throat (27.8%, 148/532), nausea (19.5%, 104/532), and vomiting (14.7%, 78/532). No serious adverse events such as gastrointestinal bleeding, perforation, cardiac or pulmonary complications occurred. Conclusion The novel portable endoscope can safely and effectively complete the diagnosis and treatment of upper gastrointestinal diseases in primary medical units, while saving the decontamination process. However, the incidence of discomfort is high during examinations. Further optimization of the operation methods is needed.