Objective：This study aims to analyze the distribution of antimicrobial resistance and the mode of aggregated multi-drug resistance in Streptococcus pneumoniae isolates in Guangzhou from 2012 to 2020. Methods：The clinical data of Streptococcus pneumoniae infection from 2012 to 2020 were collected；the preliminary identification was carried out by VITEK 2 Compact and MALDI-TOF/TOF MS，and verified again by Optoxin test and bile hemolysis test；the drug sensitivity spectrum was analyzed by VITEK 2 Compact and Kirby-Bauer method，the results were interpreted according to the CLSI2020 standard，and the data were statistically analyzed by WHONET5.6 and SPSS23.0 software. Results：A total of 1 110 strains were collected，of which 753 strains were included in the analysis，mainly from sputum samples（73.7%）. Among the infected patients，the male was on the high side（69.7%），and the age group was mainly concentrated in infants and children ≤5 years old（53.9%），the distribution of departments was mainly concentrated in ICU（51.4%）. Multidrug-resistant Streptococcus pneumoniae accounted for 75.3% of the total isolates（567/753），from 66.7%（44/66）in 2012 to 93.9%（46/49）in 2020，showing an upward trend（P < 0.001）. The resistance rate of meropenem increased from 27.9%（17/61）to 58.7%（27/46），showing an obvious increasing trend（P < 0.001）. The resistance rate of penicillin decreased from 70.0%（7/10）to 18.4%（9/49）（P < 0.001）. Cefotaxime increased from 9.1%（1/11）to 32.6%（15/46）（P=0.063），but its upward trend can not be ignored and needs further monitoring. The resistance rates of erythromycin and tetracycline have been high. Multi-drug resistance mode appeared aggregation，the most important is mode A：tetracycline-compound sulfamethoxazole-erythromycin simultaneous resistance，accounting for 27.3%（155/567），other drug resistance modes are further developed on the basis of mode A. Conclusion：Multi-drug resistance of Streptococcus pneumoniae is increasing year by year，meropenem and cefotaxime are the key monitoring drugs，penicillin resistance tends to decrease，which can be used as empirical treatment. The mode of aggregate drug resistance suggests that it is possible to experience the failure of drug use，and regional rational use of antibiotics and regular monitoring of drug resistance is a long-term task.