Caister Academic Press

Applications of Real-Time PCR in Clinical Microbiology

Authors: Andrew David Sails
Abstract: The introduction of real-time PCR assays to the clinical microbiology laboratory has led to significant improvements in the diagnosis of infectious disease. There has been an explosion of interest in this technique since its introduction and several hundred reports have been published describing applications in clinical bacteriology, parasitology and virology. There are few areas of clinical microbiology which remain unaffected by this new method. It has been particularly useful to detect slow growing or difficult to grow infectious agents. However, its greatest impact is probably its use for the quantitation of target organisms in samples. The ability to monitor the PCR reaction in real- time allows accurate quantitation of target sequence over at least six orders of magnitude. The closed-tube format which removes the need for post-amplification manipulation of the PCR products also reduces the likelihood of amplicon carryover to subsequent reactions reducing the risk of false-positives. As more laboratories begin to utilise these methods standardisation of assay protocols for use in diagnostic clinical microbiology is needed, plus participation in external quality control schemes is required to ensure quality of testing.
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