We conducted a retrospective study in 57 children (median age, 3.5 years; range, 1 month–14.5 years) with microbiologically confirmed pertussis infection over a recent 4-year period in a regional hospital in Japan. We obtained nasal swabs from all patients for Bordetella pertussis isolation as well as performed B. pertussis DNA detection using loop-mediated isothermal amplification (LAMP). Of the 57 cases, 34 (60%) were culture-positive and 57 (100%) were LAMP-positive. The frequency of each symptom was as follows: typical paroxysmal cough for over 14 days, 96% (55/57); paroxysms, 86% (49/57); posttussive vomiting, 33% (19/57); inspiratory whoop, 25% (14/57); and apnea, 12% (7/57). Hospitalization was required in 14 cases (25%), 93% (13/14) of which were aged <1 year. The proportion of patients previously immunized against diphtheria-tetanus-acellular pertussis vaccine (DTaP) was 19% (4/21) in children aged <1 year and 92% (11/12) in children aged ≥10 years. Minimum inhibitory concentrations for 6 antimicrobials (erythromycin, clarithromycin, azithromycin, minocycline, amoxicillin, and sulfamethoxazole/trimethoprim) were measured for 30 isolated strains, and all strains were susceptible to all aforementioned antimicrobials. Thus, an additional pertussis vaccination in older children is necessary, and the current macrolides-based treatment strategy is considered reasonable.