| Abstract: |
The polymerase chain reaction (PCR) is a well-established method in clinical laboratories. This chapter reviews the basics of real-time PCR, including instrumentation and detection chemistries, primer design and assay development, and quality control and quality assurance issues. Real-time PCR builds upon the basic principle of endpoint PCR. In the early cycles of PCR, reaction components are abundant and the concentration of PCR products doubles with each cycle. Primers and probes are critical for both the sensitivity and the specificity of a real-time PCR test. Quality control measures are essential to ensure the accuracy and reliability of real-time PCR assays. Multiplexing allows for the amplification and detection of more than one target in a single PCR reaction. Real-time PCR is universally faster when compared to culture-based techniques. © 2025 Elsevier B.V., All rights reserved. |
