Molecular Characterization of Multi-Drug Resistant Clinical Isolates from Healthcare Facilities

This study aimed to analyze multi-drug resistant clinical isolates collected from healthcare facilities at the molecular level.Bacterial samples from a hospital were cultured on selective media to isolate Acinetobacter, E. coli, Klebsiella sp., and Pseudomonas sp., and characterized by Gram staining and biochemical tests.DNA was extracted from overnight cultures and used for PCR amplification of the V1-V3 and V3-V9 regions of the 16S rDNA.The PCR products were confirmed on agarose gels, purified, and sequenced.Sequences were analyzed using BLAST for species identification, and accession numbers were obtained from the NCBI GenBank database.Morphological and biochemical data for pathogenic bacteria, such as Acinetobacter (K1), E. coli (K2), Klebsiella (K3), and Pseudomonas sp.(K4) were analyzed.The 16s rDNA PCR products were sequenced using a commercial automated sequencer and were submitted to GenBank and NCBI.Bacterial sequencing analysis and comparison revealed that isolates K1, K2, K3, and K4 were Acinetobacter, E. coli, Klebsiella, and Pseudomonas sp.since they had a 100% homology with the library.The species-level identification of bacteria K1, K2, K3, and K4 is based on the highest identity (90%) and their GenBank accession codes are APK1, APK2, APK3, and APK4.This study suggests that samples from tertiary hospitals contain high loads of antibiotic-resistant bacteria.A change in the antibiotic sensitivity of the identified bacteria needs to be monitored, as there is limited availability of newer medicines, and the emergence of resistant bacteria far exceeds the rate of new antibiotic development.