In view of the current situation that the nursing teaching content is lacking pertinence and cannot effectively cultivate students’ critical thinking, this paper puts forward the application analysis of two-dimensional code online testing technology in nursing teaching in colleges and universities. Through the analysis of two-dimensional code recognition technology architecture and common application fields, an intelligent nursing teaching platform for two-dimensional code online testing technology was designed. Systematically, we summarize the advantages of intelligent nursing teaching platform, use flash and 3D technology to complete the virtualization of nursing teaching scene, and use XML technology to update and save teaching resources; we further build an intelligent nursing teaching platform by using the basic plate, nursing station plate, ward plate, dispensing room plate, and case and operation review plate. The experimental results show that under the two-dimensional code online test technology, the nursing teaching method in colleges and universities has strong teaching resource processing efficiency, improves the operation level of students’ nursing technology, and effectively cultivates the academic thinking of nursing students, which is of great significance to the progress of nursing teaching.