Antibiotic resistant pathogens cannot be restricted to geographic boundaries, therefore, antibiotic resistance (ABR) is not a problem of a particular area, region or country but it is a global threat and anyone can be suffered with infections without treatment options. ABR has been recognized as complex global public health task which requires adoption of multiple approaches to control and overcome this challenge, especially for Acinetobacter sp. and Pseudomonas sp. These two organisms remained unattended even in AMR global report on surveillance by WHO. Lately, isolation of Acinetobacter and Pseudomonas with higher frequencies from hospital-acquired infections (HAIs) and emergence of highly resistant strains including XDR and even PDR evoked the interests of clinicians towards these pathogens. Consequently, in 2017, WHO also recognized these bugs as high priority threats to human health. Keeping in view the severity of the problems caused by drug resistant strains of Acinetobacter and Pseudomonas, present study targeted them by isolating from patients admitted in ICUs along with few other strains. The isolates were identified by traditional as well, highly sensitive method, matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS). So far, identification of Pseudomonas and Acinetobacter isolates by MALDI-TOF-MS has not been reported from Pakistan. This method enabled us to identify few rare species of Pseudomonas including P. balearica, P. otitidis, and P. fulva. These species are mostly reported from marine environment, food samples and ear infections. Yet the pathogenic role of these strains was not explored in this study as routine testing across the country does not identify them.