ARTICLE IN PRESS FOOD MICROBIOLOGY Food Microbiology 22 (2005) 267–268 www.elsevier.com/locate/yfmic
Book review Real-time PCR: an essential guide Edwards Kirsten, Logan Julie, Saunders Nick (Eds.), Horizon Bioscience, Wymondham, Norfolk, UK, ISBN: 0-9545232-7-X, 2004 346 pp., Hardcover. Price: GB £90 US $180. The polymerase chain reaction has become a routine technique in many research and diagnostic laboratories and real-time PCR has since its introduction in the mid1990s removed many limitations of the standard endpoint PCR. After a short introduction (Chapter 1), this book presents a very useful overview of the different real-time PCR platforms (Chapter 2) and various realtime PCR chemistries in use (Chapter 3) with their possibilities and limitations. The principle of real-time PCR and the different probe technologies are well explained. However, although huge comparative tables are part of the textbook and are very informative (e.g. a table summarizing some of the common fluorophore combinations that can be used effectively in energy transfer pairs), it is a pity that additional, higher quality figures were not included to better illustrate the various PCR machines and chemistries available (one of the figures on the fluorescent chemistries also describes colours in the legend although the graph is in black and white). In the description of the real-time PCR technique, the authors are aware that in a rapidly changing science such as molecular biology, new systems and probe designs are continuously being placed on the market, and they refer the reader to the manufacturers’ websites and other web resources for updated information. The book does not limit itself to description of the real-time PCR system itself but it has a significant added value in the description of the critical points in the performance of real-time PCR (Chapter 4). A lot of important remarks are included in the book on the practical implementation of real-time PCR that may be overlooked by the inexperienced user of real-time PCR and that may save a lot of time and money setting up a real-time PCR protocol. It also describes in a separate chapter (Chapter 5) important aspects on the use and
doi:10.1016/j.fm.2004.08.005
development of internal and external controls that are necessary to include for validation of the test results. As such it is a basic and essential manual for any instructor, student or researcher planning the introduction of realtime PCR in teaching material or experimental work. Subsequent chapters give a comprehensive overview of important real-time PCR applications such as quantitation (Chapter 6), expression analysis (Chapter 7) and mutation detection (Chapter 8 and 9). Interesting are the protocols which are explained in detail in the chapter on real-time PCR analysis of mRNA expression and which can be tried out immediately. A separate chapter describes real-time NASBA (Chapter 10), an alternative isothermal amplification system with particular application to viral diagnostics. A minor point for food microbiologists to recognize is that the final chapters of the book are dedicated to clinical microbiology applications of real-time PCR (Chapter 11) and to the diagnosis of invasive fungal infection (Chapter 12). In Chapters 10 and 11 exhaustive tables are to be found which summarize literature data on application of real-time PCR indicating the sensitivity and target gene for several target organisms (among which are bacteria and viruses). Applications that may be of interest to food microbiologists include the overview of real-time PCR for detection of Mycobacterium tuberculosis, Legionella species, verotoxin-producing E. coli and enteric parasites such as cryptosporidia and Cyclospora and real-time PCR assays for detection of antibiotic resistant strains of Campylobacter jejuni and Salmonella. Sufficient detailed information is given and the interested reader may benefit from reading it. Nevertheless, as we all know, the approach in clinical diagnostics is often a forerunner of what is happening in food diagnostics. Indeed, in the past two years, publications on the applications of real-time PCR in food microbiology have increased tremendously. This book can serve as a manual especially for those who need a basic introduction to the subject of real-time PCR, but also for those who are already familiar with the technology and are interested in the use of real-time PCR for applications such as mutation detection and
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Book review / Food Microbiology 22 (2005) 267–268
analysis of mRNA expression. It is well written and easy reading for the interested reader, although a minor remark might be that not all the abbreviations used in the book are immediately defined. Also, lay-out could have been improved (headlines and subtitles are not always easy distinguishable). In conclusion, it is a basic textbook for those laboratories that are working with real time PCR, and it can stimulate readers to develop their own applications of this technology.
M.Uyttendaele Laboratory of Food Microbiology and Food Preservation, Department of Food Technology and Nutrition, Faculty of Agricultural and Applied Biological Sciences, University of Gent, Coupure Links 653, B-9000 Gent, Belgium E-mail address:
[email protected]
