Molecular Analysis of Gram-Negative Bacterial Strains in Patients from Koya City

Abstract

This study addresses the critical issue of Gram-negative bacteria (GNB) in patients from Koya City, acknowledging their significant resistance to antibiotics and the consequent threat to public health. A total of 14 samples were collected from diverse patient demographics and sources, including urine, wounds, vaginal swabs, seminal fluid, and throat swabs. The samples were subjected to analysis through a multifaceted approach involving culturing and molecular identification. Following the isolation of bacterial samples, the 16SrRNA gene was targeted using polymerase chain reaction (PCR). Subsequently, the obtained PCR products underwent Sanger sequencing, providing a detailed analysis of the nucleotide sequences. The resulting sequences were then compared against existing prokaryotic DNA sequences in the BLAST database to determine the identity of the isolated bacteria. Out of the 14 samples, 13 yielded sequences of good quality, while one exhibited a noise sequence. The sequences were further aligned, and 11 samples demonstrated a 100% identity with previously identified bacteria such as Escherichia coli (100 %), Klebsiella pneumoniae (100 %), and Shigella flexneri (100 %) Notably, two samples displayed a >99% identity with Escherichia coli (99.5%), highlighting the diversity within the bacterial strains. This study sheds light on the prevalence and identity of Gram-negative bacteria in Koya City patients. The findings underscore the necessity of molecular techniques, such as PCR and sequencing, in clinical diagnostics. The diverse bacterial strains identified highlight the complexity of bacterial infections, emphasizing the importance of targeted approaches in addressing antibiotic resistance.