A Basic Guide to Real Time PCR in Microbial Diagnostics: Definitions, Parameters, and Everything

Real time PCR (quantitative PCR, qPCR) is now a well-established method for the detection, quantification, and typing of different microbial agents in the areas of clinical and veterinary diagnostics and food safety. Although the concept of PCR is relatively simple, there are specific issues in qPCR...

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Published in:Frontiers in microbiology Vol. 8; p. 108
Main Authors: Kralik, Petr, Ricchi, Matteo
Format: Journal Article
Language:English
Published: Switzerland Frontiers Media S.A 02.02.2017
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ISSN:1664-302X, 1664-302X
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Abstract Real time PCR (quantitative PCR, qPCR) is now a well-established method for the detection, quantification, and typing of different microbial agents in the areas of clinical and veterinary diagnostics and food safety. Although the concept of PCR is relatively simple, there are specific issues in qPCR that developers and users of this technology must bear in mind. These include the use of correct terminology and definitions, understanding of the principle of PCR, difficulties with interpretation and presentation of data, the limitations of qPCR in different areas of microbial diagnostics and parameters important for the description of qPCR performance. It is not our intention in this review to describe every single aspect of qPCR design, optimization, and validation; however, it is our hope that this basic guide will help to orient beginners and users of qPCR in the use of this powerful technique.
AbstractList Real time PCR (quantitative PCR, qPCR) is now a well-established method for the detection, quantification, and typing of different microbial agents in the areas of clinical and veterinary diagnostics and food safety. Although the concept of PCR is relatively simple, there are specific issues in qPCR that developers and users of this technology must bear in mind. These include the use of correct terminology and definitions, understanding of the principle of PCR, difficulties with interpretation and presentation of data, the limitations of qPCR in different areas of microbial diagnostics and parameters important for the description of qPCR performance. It is not our intention in this review to describe every single aspect of qPCR design, optimization, and validation; however, it is our hope that this basic guide will help to orient beginners and users of qPCR in the use of this powerful technique.Real time PCR (quantitative PCR, qPCR) is now a well-established method for the detection, quantification, and typing of different microbial agents in the areas of clinical and veterinary diagnostics and food safety. Although the concept of PCR is relatively simple, there are specific issues in qPCR that developers and users of this technology must bear in mind. These include the use of correct terminology and definitions, understanding of the principle of PCR, difficulties with interpretation and presentation of data, the limitations of qPCR in different areas of microbial diagnostics and parameters important for the description of qPCR performance. It is not our intention in this review to describe every single aspect of qPCR design, optimization, and validation; however, it is our hope that this basic guide will help to orient beginners and users of qPCR in the use of this powerful technique.
Real time PCR (quantitative PCR, qPCR) is now a well-established method for the detection, quantification, and typing of different microbial agents in the areas of clinical and veterinary diagnostics and food safety. Although the concept of PCR is relatively simple, there are specific issues in qPCR that developers and users of this technology must bear in mind. These include the use of correct terminology and definitions, understanding of the principle of PCR, difficulties with interpretation and presentation of data, the limitations of qPCR in different areas of microbial diagnostics and parameters important for the description of qPCR performance. It is not our intention in this review to describe every single aspect of qPCR design, optimization, and validation; however, it is our hope that this basic guide will help to orient beginners and users of qPCR in the use of this powerful technique.
Author Kralik, Petr
Ricchi, Matteo
AuthorAffiliation 1 Department of Food and Feed Safety, Veterinary Research Institute Brno, Czechia
2 Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna “Bruno Ubertini,” National Reference Centre for Paratuberculosis Piacenza, Italy
AuthorAffiliation_xml – name: 1 Department of Food and Feed Safety, Veterinary Research Institute Brno, Czechia
– name: 2 Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna “Bruno Ubertini,” National Reference Centre for Paratuberculosis Piacenza, Italy
Author_xml – sequence: 1
  givenname: Petr
  surname: Kralik
  fullname: Kralik, Petr
– sequence: 2
  givenname: Matteo
  surname: Ricchi
  fullname: Ricchi, Matteo
BackLink https://www.ncbi.nlm.nih.gov/pubmed/28210243$$D View this record in MEDLINE/PubMed
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Keywords accuracy
limit of detection
limit of quantification
quantitative PCR
trueness
precision
Language English
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This article was submitted to Food Microbiology, a section of the journal Frontiers in Microbiology
Edited by: Jean-christophe Augustin, Ecole Nationale Vétérinaire d'Alfort, France
Reviewed by: Silvana Vero, University of the Republic, Uruguay; Bastien Fremaux, French Pork and Pig Institute, France
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Snippet Real time PCR (quantitative PCR, qPCR) is now a well-established method for the detection, quantification, and typing of different microbial agents in the...
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SubjectTerms Microbiology
Title A Basic Guide to Real Time PCR in Microbial Diagnostics: Definitions, Parameters, and Everything
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