Environmental Applications Of Nucleic Acid Amplification Technology

From the Preface Antibody techniques have allowed us to study microorganisms in situ. However, until recently all methodology lacked the sensitivity necessary for environmental work where microorganisms are in most cases present at very low concentrations or where microbial ecosystems contain a myriad of different organisms. Gene probes have been used successfully for a variety of samples, but this method was still not sensitive enough. The next logical step was the application of the recently developed DNA amplification technique known as the polymerase chain reaction, or PCR. Since then, many laboratories around the world have adopted PCR for environmental work. Samples obtained from soils, water and air are enormously complex because they are unknown mixtures of DNA and other compounds. Thus, procedures for target DNA amplification from the environment require special attention. The PCR has allowed us to go beyond the need for culturing prior to analysis of microbial communities. It has been shown that even microorganisms that can be routinely grown in the laboratory undergo some physiological changes when exposed to the environment. One of these changes (first observed by R. Colwell and colleagues) is known as the viable-but-non-culturable state, and seems to be a common occurrence. Thus, the use of culture techniques paint only part of the picture in terms of microbial behavior under environmental conditions. The ability to amplify nucleic acids by the PCR has brought about a myriad of very ingenious modifications to the technique that can then be used to study complex ecosystems. The manner in which the PCR can be modified is only limited by the need and/or the imagination of the researcher. The first manual dedicated specifically to the analysis (by PCR) of environmental samples, Environmental Applications of Nucleic Acid Amplification Techniques presents state of the art methodology for the detection of microorganisms in soil, water, air samples, as well as the amplification of nucleic acids from fossil samples. The manual gives step-by-step procedures for the analysis of these samples. Although several publications have addressed the use of Polymerase Chain Reaction technique, very few of them have been directed toward the application of this technique to environmental samples. This book fills this gap in the literature.

This book discusses recent advances in the use of nucleic acid based biosensors and related bioanalytical assays for environmental monitoring.

Providing current information and guidance on the uses of various nucleic acid amplification technologies for clinical laboratory diagnosis, this book goes beyond the Polymerase Chain Reaction to explore a broader range of important alternative DNA/RNA amplification methods including the Ligase Chain Reaction, Q[beta] Replicase Assays and TMA. There are many examples of specific applications of these technologies, discussions of yet unresolved issues and demonstrations of the relevance of these technologies to medical research and disease diagnostics. Individual chapters cover uses of these methods in clinical situations such as detection of food pathogens, viral infections, STDs, Mycobacteria drug resistance mutations, and heritable diseases. Automation, diagnostic test evaluation, and the synthesis of artificial DNA are also discussed. This book is designed for all biomedical scientists interested in the application of molecular biology to clinical diagnosis.

Nucleic acid hybridizations: principles and strategies; Generation of nucleic acid probe molecules; Nonisotopic DNA labeling strategies; Approaches to genetic analysis: genetic disease and screening; Approaches to genetic analysis: forensics; Approaches to infectious disease diagnosis; Diagnostic nucleic acid probe technology in veterinary medicine; Diagnostic nucleic acid probes in human medicine: applications, present and future; Developments in food technology: applications and regulatory and economic considerations; Current development and applications of nucleic acid technology in the environmental sciences.

With emerging biological threats from pathogenic microorganisms and increasing environmental pollutants, it is essential to ensure the safety needs of individuals and the ecosystem are met. Modern materials science and engineering has evolved over the years to better develop devices to test abnormalities. Affordability, accessibility, and reliability of any analytical system is the prime necessity for a modern diagnostic application. Health and Environmental Applications of Biosensing Technologies: Clinical and Allied Health Science Perspective presents a detailed overview on biosensor design systems and optimal fabrication technologies to create a greater impact on various industries and help organizations break existing performance tradeoffs to deploy biosensor technologies across inter/transdisciplinary businesses. The book presents novel and emerging trends in biosensor design and healthcare applications focused on API detection, communicable/non-communicable disease diagnosis, food quality monitoring, agro-environmental analysis, bio-defense, and industrial pollutant sensing. In addition, wearable biosensors, commercial products, and safety regulations for biosensing technologies are summarized. Provides a fundamental understanding on biosensor system design, biomarkers for communicable/non-communicable diseases, and bioreceptor immobilization techniques Integrates information covering biosensing technologies for clinical diagnosis, API detection, industrial/environmental monitoring, agro-livestock healthcare, and disease control Provides information on principles, advanced trends, and approaches for wearable biosensors Covers market trends with biosensing technologies/products and their commercial challenges

In the ten years since the first publication on PCR (Saiki et al. , 1985), this in vitro method of nucleic acid replication and modification has grown to rival in popularity traditional microbiological, genetical und technical procedures for cloning, sequencing, gene detecting and related procedures. To date the PCR literature has emphasized six main areas of application: genetic mapping, detection of mutations, genetic polymorphism, transcriptional splicing and regulation, molecular virology and quantitative procedures. The overwhelming focus of quantification of DNA or RNA by PCR has been on human microbiology and oncological problems. The exquisite sensitivity of PCR gives this method the ability to detect extremely rare DNAs, mRNAs, mRNAs in small numbers of cells or in small amounts of tissue, and mRNAs expressed in mixed-cell populations. However, the exact and accurate quantification of specific nucleic acids in biological samples is in spite of numerous publications in that field still a general problem: during the peR process, an unknown initial number of target sequences are used as a template from which a large quantity of specific product can be obtained. Although the amount of product formed is easy to determine, it is difficult to deduce the initial copy number of the target molecule because the efficiency of the peR is largely unknown.

Molecular biological techniques such as DNA/RNA extraction and purification, and especially the polymerase chain reaction, PCR, are rapidly gaining interest also in related fields, such as microbiology or environmental sciences. They offer new approaches and opportunities for the determination of microbial cells, DNA and RNA from soils, roots, rhizospheres, sediments and aquatic environments. Detailed protocols for these applications are described in this manual.

Since Operation Desert Shield/Desert Storm, Gulf War veterans have expressed concerns about health effects that could be associated with their deployment and service during the war. Although similar concerns were raised after other military operations, the Gulf War deployment focused national attention on the potential, but uncertain, relationship between the presence of chemical and biological (CB) agents and other harmful agents in theater and health symptoms reported by military personnel. Strategies to Protect the Health of Deployed U.S. Forces which is one of the four two-year studies, examines the detection and tracking of exposures of deployed personnel to multiple harmful agents.