In recent years, the ST131 clone was identified as a high risk pandemic clone among Escherichia coli isolates by multilocus sequence typing (MLST) studies and has been associated with extended spectrum beta-lactamase (ESBL) production (often with CTX-M-15) and antibiotic resistance especially against fluoroquinolones. The aim of this study was to determine the rate of high risk ST131 clone in ESBL producing E.coli isolates in our region, to investigate the sensitivity of MALDI-TOF MS in the detection of ST1 31 clone, and to compare the frequency of antimicrobial resistance among ST131 and non-ST1 31 isolates. A total of 251 urinary and 50 non-urinary E.coli isolates identified in our hospital central laboratory between February 2016-February 2017 were included in the study. Real-time PCR and MALDI-TOF MS methods were used for the detection of E.coli ST131 clone. For the statistical evaluation of the rate of antibiotic resistance among isolates of ST131 and non-ST131 clones, chi-square test was used. p value under 0.05 was considered as significant. Of the 301 isolates, 110 (36.6%) and 92 (30.6%) isolates were identified as ST131 clone by real-time PCR and MALDI-TOF MS, respectively. According to real-time PCR results, 91 (36.3%) of 251 urinary isolates and 19 (38%) of 50 non-urinary isolates were found as ST1 31 clone; there was no statistically significant difference between the groups. Ciprofloxacin resistance was found to be significantly higher in ST131 isolates than the non-ST131 isolates (78.2%, n= 86 vs. 53.4%, n= 102). No statistically significant difference was determined for the other antibiotics tested. For the detection of E.coli ST131 clone; sensitivity of MALDI-TOF MS was 84%, specificity was 100% while positive predictive value was 100% and negative predictive value was 92%. In conclusion, further investigation of the high risk E.coli ST1 31 clone in our country, in which ESBL ratios and antibiotic resistance rates, especially in fluoroquinolones, are high, is important for the development of new strategies to control antibiotic resistance. MALDI-TOF MS method is particularly useful for easy and fast detection of the high risk E.coli ST131 clone.