The present study aimed at investigating the spread of serine carbapenemase-producing isolates of Pseudomonas aeruginosa recovered from some hospitals and specialized health centers in Al-Diwaniyah City, Iraq. Using morphological and molecular methods, 630 clinical samples (244 burn swabs, 163 sputa, 115 urine samples, and 108 ear swabs) were tested. The current work also explored the carbapenemase production ability of the carbapenemresistant (CR) isolates using morphological and molecular methods. A monoplex polymerase chain reaction (MPCR) technique targeting 4 different serine carbapenemase (SCMs) coding genes was followed. The study also explored the nucleotide sequences of the SCM genes. The API 20 E and Vitek 2 system showed that 100 of the isolates belonged to P. aeruginosa. With different severity of resistance by percentages of isolates to various antibiotics, only 24% of the isolates displayed resistance against carbapenem. The results of the SCM-related MPCR showed the occurrence of the bla_GES, bla_sMe, bla_KPC, and bla_OXA-48 genes in 20 (83.3%), 19 (79.1%), 11 (45.8%), 18 (75%), respectively, of the isolates. The phylogenetic study analysis revealed high, 99%, identity with NCBI-related world strains. According to the best of our knowledge, this is the first study in Iraq that detects a group of encoding genes of serine carbapenemase in carbapenem-resistant isolates.