With the wide application of parenteral nutrition (PN) and central venous catheter (CVC) in tumor patients, the relative infection led by CVC has become a common and troublesome hospital infection in clinics. During infusion, PN fluid forms a high-sugar and high-fat microenvironment around CVC, facilitating the adhesion, growth, and diffusion of pathogenic bacteria. This condition forms a bacterial biofilm (BF) consisting of one or several kinds of pathogenic bacteria. Upon formation of the film, pathogens in the BF can resist antibacterial drugs and immune cells, causing repeated infections of bacteria or fungi and endangering the lives of patients. In this article, we summarize the applications of PN, characteristics of CVC infection in tumor patients, mixed BF, and related research methods to provide reference for studies of mixed BF infection of CVC.