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Clinical Microbiology Reviews, October 2000, p. 559-570, Vol. 13, No. 4
0893-8512/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Multiplex PCR: Optimization and Application in Diagnostic Virology

Elfath M. Elnifro,1 Ahmed M. Ashshi,1 Robert J. Cooper,1 and Paul E. Klapper1,2,*

School of Medicine, The University of Manchester,1 and Clinical Virology, Central Manchester Healthcare Trust,2 Manchester, United Kingdom

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. Multiplex PCRs to detect viral, bacterial, and/or other infectious agents in one reaction tube have been described. 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.

* Corresponding author. Mailing address: Clinical Virology, 3rd Floor, Clinical Sciences Building, Manchester Royal Infirmary, Oxford Rd., Manchester M13 9WL, United Kingdom. Phone: 44 161 276 8849. Fax: 44 161 276 8840. E-mail: pklapper{at}mri5.cmht.nwest.nhs.uk.

Clinical Microbiology Reviews, October 2000, p. 559-570, Vol. 13, No. 4
0893-8512/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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