Major science and technology infrastructure is the “national weight” of China’s construction of a world science and technology power, which can sustainably promote the output of major original achievements, accelerate the integration of innovative elements and breakthrough of high-end industries. With the development of a new round of scientific and technological revolution and the advancement of economic globalization, international competition has become increasingly complex, and the development of major scientific and technological infrastructure has entered a new stage and faced new challenges. By combing and analyzing the new development situation of major science and technology infrastructure and the challenges faced by operation and management, this paper discusses the operation and management path of major science and technology infrastructure in the new era, which has reference significance for improving the construction and operation and management level of major science and technology infrastructure.