Current Issues in Molecular Biology

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Open AccessReview

Quantitative Real-Time Polymerase Chain Reaction for Tracking Microbial Gene Expression in Complex Environmental Matrices

by Vijay J. Gadkar and Martin Filion

Curr. Issues Mol. Biol. 2013, 15(2), 45-58; https://doi.org/10.21775/cimb.015.045 - 8 Apr 2013

Cited by 1 | Viewed by 666

Abstract

Environmental matrices are highly diverse in their composition and range from simple (e.g. water) to highly complex (e.g. organic soils/biosolids). Analysis of microbial gene expression from such substrates is done for variety of purposes which could range from bio-surveillance to elucidation of biological function of a target microbe. Quantitative real-time PCR (RT-qPCR) has become a technique of choice for studying such bio-processes, due to its unique ability to both detect and quantify a target transcript in real-time. Challenges in extracting inhibitor-free, structurally intact RNA, amenable for a sensitive technique like RT-qPCR, has however proved to be a major impediment in our ability to rigorously implement this highly versatile technology. Despite these 'substrate defined' limitations, many attempts have been made to implement the RT-qPCR technology. Efforts like these have given us invaluable insight into the expression status of a particular transcript and hence, the biological functioning of the microbe, specifically under natural in situ conditions. As a result, it has enhanced our understanding of the role and diversity of many microbial populations which, previously was not possible using conventional molecular approaches. In this article, we have sought to summarize such technical problems faced by molecular environmental microbiologist and solutions developed to mitigate those challenges. Full article

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Open AccessReview

Real-Time PCR in Food Science: PCR Diagnostics

by David Rodríguez-Lázaro, Nigel Cook and Marta Hernández

Curr. Issues Mol. Biol. 2013, 15(2), 39-44; https://doi.org/10.21775/cimb.015.039 - 19 Mar 2013

Cited by 36 | Viewed by 1642

Abstract

A principal consumer demand is a guarantee of the safety and quality of food. The presence of foodborne pathogens and their potential hazard, the use of genetically modified organisms (GMOs) in food production, and the correct labelling in foods suitable for vegetarians are among the subjects where society demands total transparency. The application of controls within the quality assessment programmes of the food industry is a way to satisfy these demands, and is necessary to ensure efficient analytical methodologies are possessed and correctly applied by the Food Sector. The use of real-time PCR has become a promising alternative approach in food diagnostics. It possesses a number of advantages over conventional culturing approaches, including rapidity, excellent analytical sensitivity and selectivity, and potential for quantification. However, the use of expensive equipment and reagents, the need for qualified personnel, and the lack of standardized protocols are impairing its practical implementation for food monitoring and control. Full article

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Open AccessReview

Real-Time PCR in Food Science: Introduction

by David Rodríguez-Lázaro and Marta Hernández

Curr. Issues Mol. Biol. 2013, 15(2), 25-38; https://doi.org/10.21775/cimb.015.025 - 19 Mar 2013

Cited by 25 | Viewed by 1856

Abstract

Food safety and quality control programmes are increasingly applied throughout the production food chain in order to guarantee added value products as well as to minimize the risk of infection for the consumer. The development of real-time PCR has represented one of the most significant advances in food diagnostics as it provides rapid, reliable and quantitative results. These aspects become increasingly important for the agricultural and food industry. Different strategies for real-time PCR diagnostics have been developed including unspecific detection independent of the target sequence using fluorescent dyes such as SYBR Green, or by sequence-specific fluorescent oligonucleotide probes such as TaqMan probes or molecular beacons. Full article

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Curr. Issues Mol. Biol., Volume 15, Issue 2 (July 2013) – 3 articles

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