Genetic Analysis of Spore-Forming Pathogenic Bacteria Using PCR in Clinical Samples from the Republican Hospital in Kirkuk-Iraq

Abstract

Background: Spore-forming bacteria such as Bacillus and Clostridium are well-known for their ability to endure extreme conditions and are significant contributors to hospital-acquired infections. Their spores enable them to resist heat, disinfectants, and environmental stressors. Detecting sporulation-related genes, especially spo0A, is essential for understanding their survival and pathogenicity in clinical environments. Objective: To detect and analyze spore-forming pathogenic bacteria in wound samples from hospitalized patients using molecular techniques, with an emphasis on identifying the spo0A gene as a biomarker for sporulation potential. Methods: A total of 300 wound swabs were collected from post-surgical patients at Kirkuk General Hospital (2024–2025). Samples were cultured on chocolate, blood, and MacConkey agar at 37°C. Isolates were identified using Gram staining, biochemical tests, and sporulation assessments. DNA was extracted using QIAamp kits. PCR was performed using primers specific to 16S rRNA and spo0A genes. Amplified products were visualized using agarose gel electrophoresis with ethidium bromide staining. Identified species included E. coli, Enterobacter cloacae, S. aureus, P. aeruginosa, B. subtilis, and C. perfringens. Statistical Analysis Used: The study utilized descriptive statistics, primarily percentages, to interpret the distribution of bacterial species and infection rates. No inferential statistical tests were used. Results: Of the 300 samples, 150 (50%) were positive for bacterial infection. Only B. subtilis and C. perfringens tested positive for the spo0A gene. PCR successfully identified spore-forming strains with high accuracy. Conclusion: PCR proved to be a sensitive and efficient method for detecting spore-forming genes. Understanding bacterial sporulation is vital for enhancing infection control and clinical diagnostics in healthcare environments. By identifying spore-forming bacteria quickly and correctly, PCR can improve infection control procedures in hospitals by enabling early diagnosis and prompt preventive actions.