Applications in Clinical Microbiology
Andrew D. Sails
from: Real-Time PCR: Advanced Technologies and Applications (Edited by: Nick A. Saunders and Martin A. Lee). Caister Academic Press, U.K. (2013)
The introduction of real-time PCR technology to diagnostic clinical microbiology laboratories has led to significant improvements in the diagnosis of infectious disease. It has been particularly useful to detect slow growing or difficult to grow infectious agents therefore much of its initial impact was in diagnostic virology. However, in more recent years real-time PCR-based methods have been introduced in diagnostic bacteriology, mycology and parasitology and there are few areas of clinical microbiology which remain unaffected by real-time PCR methodologies. One area where it has had great impact is its use for quantitation of viral pathogens. The ability to monitor the PCR reaction in real-time allows accurate quantitation of target sequence over at least six orders of magnitude. In addition, the closed-tube format removes the need for post-amplification manipulation of the PCR products also reducing the likelihood of amplicon carryover to subsequent reactions reducing the risk of false-positives. The inherent sensitivity of real-time PCR means that contamination between samples and from previously amplified product can lead to false positive results. Therefore diagnostic labs utilising real-time PCR methods have to strictly adhere to good laboratory practice to reduce the likelihood of cross contamination. In addition individual laboratories must ensure quality of diagnostic testing by participating in external quality assurance schemes read more ...