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

被引:640
作者
Kralik, Petr [1 ]
Ricchi, Matteo [2 ]
机构
[1] Vet Res Inst, Dept Food & Feed Safety, Brno, Czech Republic
[2] Ist Zooprofilatt Sperimentale Lombardiae & Emilia, Natl Reference Ctr ParaTB, Piacenza, Italy
关键词
quantitative PCR; limit of detection; limit of quantification; accuracy; precision; trueness; AVIUM SUBSP PARATUBERCULOSIS; ENZYMATIC AMPLIFICATION; QUANTIFICATION; DNA; VALIDATION; BACTERIA; MILK; RNA;
D O I
10.3389/fmicb.2017.00108
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
收藏
页数:9
相关论文
共 42 条
[1]  
[Anonymous], 2015, JRC TECHNICAL REPORT
[2]  
[Anonymous], 2015, GUIDELINES VALIDATIO
[3]  
[Anonymous], 2013, TERRESTRIAL MANUAL
[4]  
[Anonymous], 2012, METHODS COMMITTEE GU
[5]  
[Anonymous], 2011, 16140 ISO
[6]  
[Anonymous], 1994, Accuracy (trueness and precision) of measurement methods and results - Part 1: General principies and defmitions
[7]  
[Anonymous], PROTOCOL VALIDATION
[8]  
Anonymous R., 2014, OIE VALIDATION GUIDE
[9]  
ARMBRUSTER DA, 2008, CLIN BIOCHEM REV, V29, DOI DOI 10.3390/S20236922
[10]   A comparison of nucleic acid amplification techniques for the assessment of bacterial viability [J].
Birch, L ;
Dawson, CE ;
Cornett, JH ;
Keer, JT .
LETTERS IN APPLIED MICROBIOLOGY, 2001, 33 (04) :296-301