Molecular and computational analysis of conserved outer membranes (oprI and oprL) in MDR clinical isolates of Pseudomonas aeruginosa as potential vaccine targets from Lahore, Pakistan

Pseudomonas aeruginosa is a multidrug-resistant pathogen, highly prevalent in ICU patients and is a rising concern globally. Multidrug resistance (MDR) and virulence of P. aeruginosa has been a major reason linked to the high rates of mortality in both developed and non-developed countries. The study focused specifically on main structural outer membrane proteins mainly oprI and oprL due to their significant role on both virulence and MDR by assessing their MDR from the isolates collected from various healthcare settings in Lahore, Pakistan. A total of 65 isolated were collected from different tertiary care hospitals of Lahore, Punjab. All isolates were identified and characterized using conventional methods. MDR patterns were assessed through disk diffusion method in accordance with CLSI 2024 guidelines. Detection of virulence genes (oprI and oprL) was performed using PCR amplification. Statistical and in silico analysis was performed to analyse the prevalence of P. aeruginosa and identification of potential vaccine targets provide alternative treatment strategies against infections caused by it. Out of all 65 isolates, total of 52 isolates was reportedly positive with P. aeruginosa and showed 100% resistance to β-lactams with high susceptibility to cephalosporins and fluoroquinolones. Multidrug resistance was reported in 70% isolates of P. aeruginosa. All isolates (100%) harbored the oprL gene while the oprI gene was detected in 85.5% of isolates. In silico characterization revealed strong binding B and T cell epitopes with 100% conservancy as potential multi-epitope candidates that warrant further in vivo and in vitro validation to overcome deadly infections caused by P. aeruginosa. In conclusion, MDR and virulence gene characterization by both molecular and computational methods will aid in developing multi-epitope vaccine candidate that can be tested in vitro and in vivo to overcome MDR and rising mortality rates in Pakistan.