Real-time PCR in clinical microbiology: verification, validation, and contamination control

Real-time PCR is increasingly utilized in the clinical microbiology laboratory setting. Availability of a single set of guidelines for real-time PCR testing which incorporate all the necessary verification and validation recommendations set forth by accreditation agencies would be beneficial to laboratories acquiring this new technology. Presently, the challenges to the clinical microbiology laboratory for introducing real-time PCR test systems include what verification experiments are required and how many and which types of specimens should be tested. In addition, based on the regulations, the laboratory must determine what constitutes an acceptable quality control program. Contamination of specimens or samples in real-time PCR testing also needs to be addressed. Although real-time PCR test systems are closed systems, contamination from specimen-to-specimen or from previously-amplified product can lead to false-positive results causing delays in clinical diagnosis, along with wasting valuable time and resources.