Official websites use. Share sensitive information only on official, secure websites. Corresponding author. Phone: 44 Fax: 44 E-mail: pklapper mri5. PCR has revolutionized the field of infectious disease diagnosis. To overcome the inherent disadvantage of cost and to improve the diagnostic capacity of the test, multiplex PCR, a variant of the test in which more than one target sequence is amplified using more than one pair of primers, has been developed.
Early studies highlighted the obstacles that can jeopardize the production of sensitive and specific multiplex assays, but more recent studies have provided systematic protocols and technical improvements for simple test design. The most useful of these are the empirical choice of oligonucleotide primers and the use of hot start-based PCR methodology.
These advances along with others to enhance sensitivity and specificity and to facilitate automation have resulted in the appearance of numerous publications regarding the application of multiplex PCR in the diagnosis of infectious agents, especially those which target viral nucleic acids. This article reviews the principles, optimization, and application of multiplex PCR for the detection of viruses of clinical and epidemiological importance.
During the past decade, advances in PCR technology and other DNA signal and target amplification techniques have resulted in these molecular diagnostics becoming key procedures 4 , , Such techniques are conceptually simple, highly specific, sensitive, and amenable to full automation 54 , The most mature of these technologies, PCR, is in one variant or another now common in research laboratories and is used increasingly in routine diagnostic laboratory settings and undergraduate and high-school teaching 32 , 38 , 40 , In diagnostic laboratories the use of PCR is limited by cost and sometimes the availability of adequate test sample volume.
To overcome these shortcomings and also to increase the diagnostic capacity of PCR, a variant termed multiplex PCR has been described. In multiplex PCR more than one target sequence can be amplified by including more than one pair of primers in the reaction. Multiplex PCR has the potential to produce considerable savings of time and effort within the laboratory without compromising test utility.
