Background We aimed to investigate the characteristics and outcomes in septic patients with positive cultures compared with negative cultures. Methods This was a sub-analysis of a cross-section survey study including 44 hospitals in mainland China from December 1, 2015 to January, 31,2016. All septic patients diagnosed according to sepsis-1 criteria admitted to participating ICU, with cultures from any suspected sites of infection obtained at admission. Demographic information, patient source, clinical feature, laboratory findings and severity of sepsis were comprehensively collected for comparison the characteristics between culture positive and negative patients. Univariable and multivariable regression were used to find the factors associated with positive culture and propensity score matching (PSM) was performed to compare the mortality. Results A total of 1919 patients were enrolled in this study with 962 (50.1%) positive-culture and 975 (49.9%) negative-culture. Positive culture was associated with older age, male gender, transferred patients, comorbidities including coronary heart decease and solid tumor, prolonged priori ICU stay, hospital-acquired infection, pneumonia and bloodstream infection, also with higher body temperature, heart rate, procalcitonin levels and with higher APACHE II and SOFA score at admission. Negative culture was associated with prior antibiotic exposure and community-acquired infections. Patients with positive culture had an insignificantly higher mortality (37.5% vs. 34.5, P = 0.165). However, PSM revealed decreased mortality in patients with positive culture (38.2% vs. 48.2%, P = 0.012), and a significantly higher proportion of patients received targeted antibiotic treatment in survivors (37.0% vs. 46.2%, P = 0.031). Conclusions A myriad of factors may affect the results of culture. Septic patients were likely to benefit from positive cultures in mortality possibly due to the higher proportion to receive targeted antibiotic treatment, and obtaining positive culture as possible should be strongly encouraged.