DEVELOPMENTS IN QUANTITATIVE PCR

In recent years the growing interest in quantitative applications of t he polymerase chain reaction (PCR) has favoured the development of a l arge number of assay procedures suitable for this purpose. In this pap er we review some basic principles of quantitative PCR and in particul ar the role of reference materials and calibrators and the different s trategies adopted for nucleic acid quantification. We focus on two met hodological approaches for quantitative PCR in this review: competitiv e PCR and real-time quantitative PCR based on the use of fluorogenic p robes. The first is one of the most common methods of quantitative PCR and we discuss the structure of the competitors and the various assay procedures. The second section is dedicated to a recent promising tec hnology for quantitative PCR in which the use of fluorogenic probes an d dedicated instrumentation allows the development of homogeneous meth ods. Assay performance of these methods in terms of practicability and reliability indicates that these kinds of technologies will have a wi despread use in the clinical laboratory in the near future.