This study aims to isolate clinical adenovirus among hospitalized children in Seremban, Malaysia. After that, determination of the serotype and genetic relatedness of isolated adenovirus through phylogenetic analysis with the published database of human adenovirus from GenBank will be carried out. 26 HAdV-positive retrospective respiratory specimens were collected from pediatric patients with respiratory distress at the Pediatric Department of Hospital Tuanku Jaafar in Seremban, Malaysia. HAdV was isolated in A549 cell culture, sequenced and analysed for HAdV typing. Based on hexon, penton, DNA polymerase, fiber, and virus-associated (VA) RNAII genes, there were 22/26 samples (84.62%) and 3/26 samples (11.54%) belonging to serotypes 7 and 3, respectively. Also, 1/26 sample (3.85%) was determined as HAdV subgroup D. In short, this study serves as a continuation to monitor the prevalence of HAdV serotypes in Malaysia and our result shows HAdV B as the most predominant circulating group of HAdV in Malaysia between 2013 – 2017.