Real-time PCR methodologies for the detection of viruses are now firmly entranced in many clinical laboratories, offering the major advantage of improved sensitivity over more traditional methods. The application of this technology has been most successful in those areas where conventional virological techniques did not exist or were inadequate. Its introduction into the virology laboratory has led to the recognition and improved diagnostics of several viruses that may not be isolated, allowed for quantification of viral loads during the infection process and the subsequent monitoring of antiviral drug therapy, as well as characterization of mutations in viral genes that are responsible for drug resistance.
The number of commercially available assays is increasing, but many laboratories still design, develop and validate assays in house. As these in-house, realtime PCR methods have evolved there has been an increasing awareness of the need for standardized reagents and quality control programs in order to produce clinically significant results. In addition, the targeted nature of PCR brings with it some inherent limitations that are difficult to control, particularly in virology, where the heterogeneous nature of the viral genome may lead to difficulties in assay design and performance. An awareness of these issues will ultimately result in a better understanding of this new technology, and enable us to fully explore its potential as a modern diagnostic tool.
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