Objective:To assess the magnitude and antimicrobial susceptibility patterns of Streptococcus pneumoniae isolates from various clinical specimens. Methods:A record based on retrospective study was conducted at Gondar University Teaching Hospital from September 2007 to January 2012. All patients who visited Gondar University Hospital and provided clinical specimens (body fluids, discharge, swab and blood) for routine bacteriological culturing and antimicrobial susceptibility testing were taken for analysis. Clinical specimens were processed for bacterial culture according to the standard procedures. Antimicrobial susceptibility test for isolated organisms was done using agar disk diffusion method. The data were entered and analyzed using SPSS software version 16 package. Results: One hundred and fifty three Streptococcus pneumoniae were isolated from patients who visited Gondar University Teaching Hospital bacteriology laboratory for culture. Majority of the pneumococcal isolates were from inpatients [111(72.5%)], and 74(48.4%) were from body fluids. Out of the total isolates, 93(61%) were found to be resistant to at least one antibiotic used for susceptibility testing. Forty eight (43.2%) of the isolates were multi-drug resistant (resistant to two or more drugs). The resistance rate noted for both ciprofloxacin 17(11.1%) and ceftriaxone 15(9.8%) were alarming. Conclusions: High proportions of the isolates tend to be increasingly resistant to the commonly prescribed drugs. The recommended drug of choice like ciprofloxacin and ceftriaxone were found to be less susceptible in the study area. Based on the findings, we therefore recommend that antimicrobial agents should be inspected for acceptable activity before they are prescribed and administered empirically. Further study with a better design and survey of antimicrobial susceptibility at large scale shoule be performed to draw advanced information.