Real-Time PCR

"... written by international authors expert in specific technical principles and applications ... a useful compendium of basic and advanced applications for laboratory scientists. It is an ideal introductory textbook and will serve as a practical handbook in laboratories where the technology is employed."

from Christopher J. McIver in Australian J. Med. Sci. 2009. 30(2): 59-60

Further reading: Real-Time PCR: Current Technology and Applications

Labels: book review, books, PCR, qPCR, real-time PCR, rtpcr

Real-Time PCR: Current Technology and Applications

Publisher: Caister Academic Press
Editors: Julie Logan, Kirstin Edwards and Nick Saunders
ISBN: 978-1-904455-39-4
"provides a dual focus by aiming, in the early chapters, to provide both the theory and practicalities of this diverse and superficially simple technology, counter-balancing this in the later chapters with real-world applications, covering infectious diseases, biodefence, molecular haplotyping and food standards." from Microbiology Today

"a reference work that should be found both in university libraries and on the shelves of experienced applications specialists." review from Microbiology Today

Further reading: Real-Time PCR

Labels: book review, qPCR, Quantitative PCR, real-time PCR

A comparison of a range of commercially available qPCR machines or real-time PCR machines lists various features to help you decide which qPCR instrument is most suitable for your needs.

• Applied Biosystems: ABI 7300, ABI 7500, ABI 7500 Fast, ABI 7900 Fast HT with automation accessory, ABI StepOne
• Roche: LightCycler 480, LightCycler 1.5, LightCycler 2.0
• Stratagene: Mx4000, Mx3000P, Mx3005P
• Cepheid: SmartCycler
• Corbett: Rotor-Gene 6000
• Eppendorf: Mastercycler ep realplex
• BioRad: MiniOpticon, MyiQ, Opticon2, Chromo4, iQ5

read more ...

Bibliography:

1. Real-Time PCR: Current Technology and Applications
2. Real-Time PCR in Microbiology: From Diagnosis to Characterization
3. PCR Troubleshooting: The Essential Guide
4. PCR Books

Labels: PCR, qPCR, real-time PCR

One of the most important recent innovations in Clinical Microbiology is the introduction of molecular methods for the diagnosis of microbial infection. The use of real-time PCR assays in the clinical microbiology laboratory has led to significant improvements in the diagnosis of infectious disease. Real-time PCR is particularly useful for the detection of slow growing or difficult to grow
infectious agents and also for the quantitation of specific organisms in samples.

Andrew Sails has written a comprehensive review on the clinical applications of Real-Time PCR in the clinical microbiology laboratory. A recently published book review described his chapter as follows: "The most comprehensive chapter is Chapter 13 Application in Clinical Microbiology, in which large numbers of microorganisms are mentioned that can be readily identified by RT-PCR." Read more of this book review at Curr. Iss. Mol. Biol.

Further reading:

• Real-Time PCR: Current Technology and Applications
• Real-Time PCR in Microbiology: From Diagnosis to Characterization

Labels: book review, diagnosis, PCR, real-time PCR

from Christina Egan, Nick M. Cirino and Kimberlee A. Musser in Real-Time PCR: Current Technology and Applications

With the public's reawakened concern regarding use of biological agents as weapons, the rapid detection, discrimination, and identification of pathogenic organisms and toxins has become a priority for state and federal government agencies. High confidence, cost effective, and near real-time diagnostic methods are essential to protecting national health security whether the target is public health, agriculture, commodities, or water supply infrastructures. While culture-based methods have been, and will likely remain, the gold standard for microbiological diagnostics, PCR-based tests offer significant advantages in sensitivity, specificity, speed and data richness that make them invaluable to diagnostic laboratories. We describe the application of real-time PCR methods in biodefense and discuss the use of real-time PCR in biodefense in terms of general workflow and processing considerations, clinical diagnostic applications, environmental diagnostic applications, and multiplex screening. Real-time PCR assays can be either quantitative (qPCR) or qualitative, depending on whether a standard curve is included with the analytical run. Most diagnostic and biodefense applications utilise the qualitative nature of real-time PCR as a detection platform. We consider the future uses and anticipated advances in real-time PCR applications as related to biodefense.

Further reading: Real-Time PCR: Current Technology and Applications

Labels: biodefense, PCR, qPCR, real-time PCR

A number of book reviews on microbiology books have been published recently. Extracts provided below:

BACILLUS: CELLULAR AND MOLECULAR BIOLOGY
"The quality of the chapters is uniformly high. Together they provide a review of significant progress toward a better and deeper understanding of the physical structure and molecular biological organization and function in Bacillus subtilis. As a consequence, a truly intimate grasp of this bacterium is achieved." from The Quarterly Review of Biology
Further information: BACILLUS

REAL-TIME PCR IN MICROBIOLOGY: FROM DIAGNOSIS TO CHARACTERIZATION
"... enjoyable and easy to read ... well-referenced and provide an easy to follow explanation, especially the chapter on fluorogenic chemistry which also has clear illustrations. The table of design rules for oligonucleotide and probes provided a good summary in an easy-to-read format. ... an interesting insight into the thoughts of the current leaders in their fields. This book shows what a versatile and sensitive tool real-time PCR can be and the many uses it can be put too. In my opinion this book would be useful for the novice because of the background information it provides." from Microbiology Today
Further information: REAL-TIME PCR

PSEUDOMONAS: GENOMICS AND MOLECULAR BIOLOGY
"... the contribution by Fajardo & Martinez on antibiotic resistance provides a wealth of background information on this clinically relevant topic ... the contributions by Haussler and Klockgether et al. give insight into recent progress made in the areas of small colony variants and genome diversity of P. aeruginosa, respectively. Three other chapters provide useful complementary information ... Institutions with an interest in Pseudomonas research should consider acquiring this book." from Microbiology Today
Further information: PSEUDOMONAS

MOLECULAR ORAL MICROBIOLOGY
"... a broad and comprehensive text covering the molecular aspects of dental caries and periodontal disease. Accompanying each of the fully referenced chapters are detailed illustrations and the inclusion of high-quality colour equivalents of these figures at the end of the book is welcome. ... of value to all microbiologists with an interest in molecular microbiology ... excellent overview of current methodologies ... appeal to dental undergraduates and postgraduates." from Microbiology Today
Further information: ORAL MICROBIOLOGY

EPIGENETICS
"... a significant collection of articles, relating to various aspects of epigenetics. The text is clear and concise and all reports include accurate data and figures. ... will assist researchers in the field and provides an important introductory reference for scientists that want to embark on such research. It is highly recommended for personal and institutional purchase." from Microbiology Today
Further information: EPIGENETICS

Labels: bacillus, book review, epigenetics, oral microbiology, Pseudomonas, real-time PCR

Following on from their highly acclaimed book "Real-Time PCR: An Essential Guide" published in 2004, Edwards, Logan and Saunders have recently completed a new, fully updated and expanded manual entitled Real-Time PCR: Current Technology and Applications. Publication is planned for January 2009 but the publishers hope to have copies available from October 2008.

Further details: Real-Time PCR: Current Technology and Applications

Labels: PCR, real-time PCR

A number of published reviews on Real-Time PCR have recently been made available as "open-access" papers. These timely and authoritative reviews are written by experts in the field and can be downloaded at: PCR Papers


Further info: PCR Papers

Labels: PCR, real-time PCR

Molecular diagnostic methods are currently being used for the early detection of many viral, bacterial, parasitic and fungal infections. Due to their high specificity and sensitivity, these methods will be inserted in the routine of the clinical laboratories to complement information provided by more conventional methods and above all, to help in the diagnosis of dubious cases. The design of specific probes, coupled to the Polymerase Chain Reaction (PCR) technique provides the required specificity and sensitivity to identify fungal species in a short time. Multicopy genes such as those of ribosomal origin are preferred by many researchers to achieve well defined results. However, because they may generate false positive results, other researchers have looked at single-copy genes of high specificity. This technology is being applied to the identification of frequently found causative agents of systemic mycoses such as Candida albicans, Aspergillus fumigatus and other species within these genera, or agents of less frequent mycoses produced by Coccidioides immitis, Cryptococcus neoformans, Histoplasma capsulatum, Paracoccidioides brasiliensis, among others.

Further reading: Pathogenic Fungi: Insights in Molecular Biology

Labels: diagnosis, fungi, mycology, PCR, real-time PCR