Streptococcus pneumoniae is one of the leading causes of death worldwide. It disseminates through colonizers and causes serious infections. Aims of this study are to determine pneumococcal carriage rate, resistance, serotype distribution, and coverage of pneumococcal conjugate vaccines from children attending day care centers from Irbid and Madaba in Jordan. Nasopharyngeal swabs were collected from day care centers (DCCs) of healthy Jordanian children 2-4 years of age from four regions of Madaba (n = 596), and from eastern Irbid (n = 423). Swabs were cultivated on Columbia blood agar base supplemented with 5% sheep blood and incubated for 18-24 hours at 37˚C with 5% CO 2. Alpha-hemolytic isolates were tested for optochin sensitivity and bile solubility for identification. Isolates were analyzed for antimicrobial susceptibility by the Vitek2 system and E-test (BioMé rieux). Serotyping was performed using the Neufeld Quellung method. A total of 341 pneumococcal strains were isolated from 1019 nasopharyngeal (NP) samples of healthy children attending DCCs for two winter seasons from 2017-2019. Carriage rate in eastern Irbid for both seasons was 29.6% and for Madaba 37.9%. Resistance rates for Irbid and Madaba, respectively, were as follows: Penicillin (86.3%; 94.4%), erythromycin (57.0%; 78.2%), clindamycin (30.8%; 47.2%), trimethoprim-sulfamethoxazole (68.6%; 86.6%), and tetracycline (45.7%; 51.9%). Predominant serotypes for Irbid were 19F (20.8%), 23F (12.0%), 6A (10.4%), and 6B (9.6%); whereas for Madaba were 19F (24.5%), 14 (7.4%), 6A (6.9%) and 23F (6.5%). Serotype coverage of the thirteen valent pneumococcal conjugate vaccine (PCV13) was about 65% for both regions. Over 96% of isolates with PCV13 serotypes in this study were resistant to penicillin with the exception of serotypes 3 and 5. As a conclusion resistance and carriage rates among the age group 2 to 4 years reached an alarming rate especially among vaccine types, which can be controlled by pneumococcal conjugate vaccination strategies.