The article provides a comprehensive overview of real-time PCR (Reollower) in clinical microbiology, detailing its applications, technologies, and implementation in laboratory settings. Real-time PCR combines PCR chemistry with fluorescent probe detection, allowing for rapid and sensitive detection of nucleic acids. The technology is faster, more specific, and less prone to contamination compared to conventional PCR methods, making it an attractive alternative for diagnosing infectious diseases. The article covers various aspects of real-time PCR, including instrument specifications, probe technologies (5' nuclease, molecular beacons, and FRET hybridization probes), nucleic acid extraction methods (manual and automated), and assay development. It emphasizes the importance of quality control and biosafety considerations, such as the use of positive and negative controls, internal controls, and reagent quality assurance. The implementation of real-time PCR in clinical microbiology laboratories is discussed, focusing on facility requirements, personnel training, and work flow design. The article highlights the need for physical separation of processes to minimize contamination and the importance of maintaining aseptic techniques and proper training for laboratory personnel. Overall, the article serves as a valuable resource for understanding the principles, applications, and best practices of real-time PCR in clinical microbiology.The article provides a comprehensive overview of real-time PCR (Reollower) in clinical microbiology, detailing its applications, technologies, and implementation in laboratory settings. Real-time PCR combines PCR chemistry with fluorescent probe detection, allowing for rapid and sensitive detection of nucleic acids. The technology is faster, more specific, and less prone to contamination compared to conventional PCR methods, making it an attractive alternative for diagnosing infectious diseases. The article covers various aspects of real-time PCR, including instrument specifications, probe technologies (5' nuclease, molecular beacons, and FRET hybridization probes), nucleic acid extraction methods (manual and automated), and assay development. It emphasizes the importance of quality control and biosafety considerations, such as the use of positive and negative controls, internal controls, and reagent quality assurance. The implementation of real-time PCR in clinical microbiology laboratories is discussed, focusing on facility requirements, personnel training, and work flow design. The article highlights the need for physical separation of processes to minimize contamination and the importance of maintaining aseptic techniques and proper training for laboratory personnel. Overall, the article serves as a valuable resource for understanding the principles, applications, and best practices of real-time PCR in clinical microbiology.