Purpose Blood is one of the most important connective tissues of human body. Bloodstream infection can range from inapparent bacteremia till fulminant septic shock with high mortality. Presence of microbes in blood whether continuously, intermittently, or transiently is a grave risk to every organ of body. Culture of blood is a vital tool to diagnose such infections. Drug susceptibility patterns help in rationalizing therapy. Objective The aim of the study is to perform bacteriological analysis and assess drug sensitivity patterns of blood culture isolates and compare in light of other associated variables. Design Retrospective observational study was conducted from January 2009 to December 2013 at a tertiary care hospital at Shillong, India. Blood samples were collected with aseptic guidelines and cultured for 7 days. Growths were identified by standard biochemical tests and subjected to sensitivity testing according to Modified Kirby Bauer disk diffusion method. Data for source of blood collection and duration of incubation were noted and compared. Results A total of 658 (11.2%) pathogens were isolated from 5,867 bacteremia-suspected patient blood specimens. Contamination was observed at the rate of 1.21%. Gram-negative organisms were the predominant pathogens recovered, Klebsiella pneumoniae being the most common. No significant difference was observed between the number of organisms isolated within or beyond 48 hours. Acinetobacter baumannii and K. pneumoniae have significantly higher chances (p < 0.05) of isolation from central line catheters compared with peripheral venipuncture. Conclusion Successful treatment of sepsis depends on early diagnosis and proper antimicrobial therapy. Local knowledge of bacteriological profile and antimicrobial sensitivity patterns helps rationalize empiric treatment strategies.