Edited by Anthony A. Killeen (Methods in Molecular Medicine), 508 pp, with illus, Totowa, NJ, Human Press, 2000.
Dr Killeen states in his preface that this book is “intended for clinical laboratory use by medical technologists and pathologists.” With this in mind, we were favorably impressed by the choice of protocols included and the ease with which they could be followed. The selections were made, according to Dr Killeen, to include those protocols determined to be used frequently or to be important for diagnostic purposes. Each of the protocols follows a logical pattern that continues from one chapter to the next.
Molecular biology is poised to revolutionize laboratory medicine, just as it has dramatically changed biomedical research. Because of its unique position at the interface between the bench and the bedside, laboratory medicine is often the first clinical discipline to apply scientific advances made by the research community. Thus, it is not surprising that the “molecular revolution” is already beginning to have a practical impact in laboratory diagnosis, even as graduate and postgraduate medical education curricula lag behind the rapid pace of research in this field.
Health care professionals in the very near future will be expected to use diagnostic profiles including complex molecular information side by side with routine clinical pathology tests. However, the task of developing reliable clinical tests from molecular methods used in research is far from straightforward. In basic and even translational research, the goal of using a particular technique is to answer a specific question in one's experimental setting. Thus, protocols are routinely modified from laboratory to laboratory and adapted to different experimental models, equipment, facilities, and operators' preferences. Because of these variables, replicating even published molecular protocols from one research laboratory to another is notoriously difficult.
The volume presents a carefully selected body of molecular pathology methods that are of wide interest to diverse medical specialties, from oncology to cardiology to infectious diseases. These include some of the most technically challenging but generally useful applications of molecular pathology, such as the extraction of usable RNA from clinical specimens. Complex techniques utilizing cell enrichment followed by reverse-transcriptase polymerase chain reaction are described in great detail. The high sensitivity of these techniques is advantageous for the detection of very rare targets, such as cancer cells in the bloodstream. At the same time, this high sensitivity increases the false-positive rate, and thus the need for stringent standardization.
A summary of the theory involved in each protocol is given in the chapter introductions, as well as a brief overview of the importance of the assay as a diagnostic tool. Many of the authors explain why they have chosen the method they discuss and why it works well in the laboratory. Reading these introductions reinforces the importance of molecular techniques in today's medicine. These techniques are used not only in diagnosis but also in following the progression and status of a disease pursuant to treatment, as in the cases of posttransplant lymphoproliferative disease and minimum reacting dose.
Each method is described completely, and charts and graphs are included when appropriate. In some cases, suggestions of worksheet design also are provided. In virtually every chapter, readers are cautioned that they must avoid contamination; because this is a constant concern in a molecular laboratory, it cannot be overstressed. Many helpful hints are given, including discussions on using different types of pipette tips, using cap covers when uncapping tubes, changing gloves often, having only one tube open at a time, and having separate processing areas for polymerase chain reaction steps.
Each of the authors shares difficulties that might be encountered in using the method he or she discusses and also some of the options available at different steps.
Each author includes a list of references for those seeking articles and papers pertaining to subjects discussed in the chapters. Reviewing these references is a good way to check on recent updates regarding diagnostic techniques as well as the history behind their use.
All methods include a section on interpretation and analysis. Many of these analyses are complex and require a separate section, whereas others are more straightforward and are explained in the introduction.
All in all, this is a very valuable reference text that belongs on the bookshelf of any clinical laboratory that has made a commitment to “going molecular.”