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Multiplex PCR: Optimization and Application in Diagnostic Virology

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Elnifro, Elfath M., Ashshi, Ahmed M., Cooper, Robert J., Klapper, Paul E.·2000·Clinical Microbiology Reviews

DOI 10.1128/cmr.13.4.559-570.2000PMC PMC88949

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Abstract

SUMMARY: 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.

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Authors: Elnifro, E. M., Ashshi, A. M., Cooper, R. J., & Klapper, P. E.
Affiliations: University of Manchester (GB)
School of Medicine, The University of Manchester,1 and
ORCID: Klapper, Paul E.
Venue: Clinical Microbiology Reviews
Publisher: American Society for Microbiology
Published: 2000-10
Volume: 13
Issue: 4
Pages: 559-570
ISSN: 0893-8512 (ISSN-L)
1098-6618
DOI: 10.1128/cmr.13.4.559-570.2000
PMC: PMC88949
OpenAlex: W2137089963
Cited by: 964
References: 126
Citations by year: 2026: 54 · 2025: 58 · 2024: 67 · 2023: 60 · 2022: 50 · 2021: 57 · 2020: 51 · 2019: 38 · 2018: 29 · 2017: 37 · 2016: 22 · 2015: 43 · 2014: 30 · 2013: 46 · 2012: 40
FWCI: 0.55
Type: journal article
Language: en
Open access: green
europepmc.org
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
Locations: Clinical Microbiology Reviews (publishedVersion)
Europe PMC (PubMed Central) (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Research Explorer (The University of Manchester) (publishedVersion)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
Physical Sciences › Engineering › Biomedical Engineering › Biosensors and Analytical Detection
Health Sciences › Medicine › Microbiology (medical) › Bacterial Identification and Susceptibility Testing
Keywords: multiplex PCR, PCR optimization, diagnostic virology, virus detection, primer design, hot-start PCR, infectious disease diagnosis, viral nucleic acids, assay sensitivity and specificity
MeSH terms: Humans, Virology, Virus Diseases, Viruses, Polymerase Chain Reaction

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Multiplex PCR: Optimization and Application in Diagnostic Virology

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