Necrotizing enterocolitis (NEC) continues to be a major cause of morbidity and mortality in low birth weight infants. Although decades of research point to a role for gut bacteria in the pathogenesis of the disease, the exact relationship between microbes and NEC has not been elucidated. In this review, we describe recent advances in the use of molecular methods to compare gut bacteria in infants with and without NEC.Our understanding of how bacteria contribute to NEC pathogenesis has been limited by the use of traditional, culture-based investigations. Recent advances in microbial ecology and DNA sequencing have made it possible to comprehensively study gut bacterial populations and to understand their physiologic importance. Several studies have identified differences in the microbiota among infants with and without NEC, but the findings have often varied across studies.To date, no single change in the gut microbiota has definitively been identified as a risk factor or cause of NEC. The findings at present suggest that NEC does not result from growth of a single causative pathogen, but rather that the disease results from a generalized disturbance of normal colonization patterns in the developing gut.