Why the need for qPCR publication guidelines?-The case for MIQE

被引:268
作者
Bustin, Stephen A. [1 ]
机构
[1] Queen Mary Univ London, Barts & London Sch Med & Dent, Blizard Inst Cell & Mol Sci, London E1 1BB, England
关键词
PCR; qPCR; DNA; RNA; Quantification; Diagnostics; REAL-TIME PCR; POLYMERASE-CHAIN-REACTION; REVERSE-TRANSCRIPTION PCR; COLORECTAL-CANCER PATIENTS; PERSISTING MEASLES-VIRUS; GENE-EXPRESSION; MESSENGER-RNA; RT-PCR; PERIPHERAL-BLOOD; QUANTITATIVE PCR;
D O I
10.1016/j.ymeth.2009.12.006
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The polymerase chain reaction (PCR) has matured from a labour- and time-intensive, low throughput qualitative gel-based technique to an easily automated, rapid, high throughput quantitative technology. Real-time quantitative PCR (qPCR) has become the benchmark technology for the detection and quantification of nucleic acids in a research, diagnostic, forensic and biotechnology setting. However, ill-assorted pre-assay conditions, poor assay design and inappropriate data analysis methodologies have resulted in the recurrent publication of data that are at best inconsistent and at worst irrelevant and even misleading. Furthermore, there is a lamentable lack of transparency of reporting, with the "Materials and Methods" sections of many publications, especially those with high impact factors, not fit for the purpose of evaluating the quality of any reported qPCR data. This poses a challenge to the integrity of the scientific literature, with serious consequences not just for basic research, but potentially calamitous implications for drug development and disease monitoring. These issues are being addressed by a set of guidelines that propose a minimum standard for the provision of information for qPCR experiments ("MIQE"). MIQE aims to restructure to-day's free-for-all OCR methods into a more consistent format that will encourage detailed auditing of experimental detail, data analysis and reporting principles. General implementation of these guidelines is an important requisite for the maturing of qPCR into a robust, accurate and reliable nucleic acid quantification technology. (C) 2009 Elsevier Inc. All rights reserved.
引用
收藏
页码:217 / 226
页数:10
相关论文
共 120 条
[1]   Absence of detectable measles virus genome sequence in blood of autistic children who have had their MMR vaccination during the routine childhood immunization schedule of UK [J].
Afzal, MA ;
Ozoemena, LC ;
O'Hare, A ;
Kidger, KA ;
Bentley, ML ;
Minor, PD .
JOURNAL OF MEDICAL VIROLOGY, 2006, 78 (05) :623-630
[2]   Model based analysis of real-time PCR data from DNA binding dye protocols [J].
Alvarez, Mariano J. ;
Vila-Ortiz, Guillermo J. ;
Salibe, Mariano C. ;
Podhajcer, Osvaldo L. ;
Pitossi, Fernando J. .
BMC BIOINFORMATICS, 2007, 8 (1)
[3]   Transcriptome Profiling Using Next-Generation Sequencing [J].
Asmann, Yan W. ;
Wallace, Michael B. ;
Thompson, E. Aubrey .
GASTROENTEROLOGY, 2008, 135 (05) :1466-1468
[4]   Increased cell-to-cell variation in gene expression in ageing mouse heart [J].
Bahar, Rumana ;
Hartmann, Claudia H. ;
Rodriguez, Karl A. ;
Denny, Ashley D. ;
Busuttil, Rita A. ;
Dolle, Martijn E. T. ;
Calder, R. Brent ;
Chisholm, Gary B. ;
Pollock, Brad H. ;
Klein, Christoph A. ;
Vijg, Jan .
NATURE, 2006, 441 (7096) :1011-1014
[5]   Gene expression profiling in single cells from the pancreatic islets of Langerhans reveals lognormal distribution of mRNA levels [J].
Bengtsson, M ;
Ståhlberg, A ;
Rorsman, P ;
Kubista, M .
GENOME RESEARCH, 2005, 15 (10) :1388-1392
[6]   Quantification of mRNA in single cells and modelling of RT-qPCR induced noise [J].
Bengtsson, Martin ;
Hemberg, Martin ;
Rorsman, Patrik ;
Stahlberg, Anders .
BMC MOLECULAR BIOLOGY, 2008, 9
[7]  
Böhlenius H, 2007, SCIENCE, V316, P367
[8]   Limitations of specific reverse-transcriptase polymerase chain reaction markers in the detection of metastases in the lymph nodes and blood of breast cancer patients [J].
Bostick, PJ ;
Chatterjee, S ;
Chi, DD ;
Huynh, KT ;
Giuliano, AE ;
Cote, R ;
Hoon, DSB .
JOURNAL OF CLINICAL ONCOLOGY, 1998, 16 (08) :2632-2640
[9]  
Burns M, 2007, EUR FOOD RES TECHNOL, V226, P7, DOI [10.1007/s00217-006-0502-y, 10.1007/s00217-007-0683-z]
[10]   Standardisation of data from real-time quantitative PCR methods - evaluation of outliers and comparison of calibration curves [J].
Burns, MJ ;
Nixon, GJ ;
Foy, CA ;
Harris, N .
BMC BIOTECHNOLOGY, 2005, 5 (1)