Clinical Applications of PCR

ISBN-13: 9780896034990 / Angielski / Twarda / 1998 / 353 str.

Lo; Y. M. Dennis Lo; Y. M. Dennis Lo

Opis

The polymerase chain reaction (PCR) is one of the most important molecular biological methods ever devised, with numerous applications to cli- cal molecular medicine. Since its description in 1985, PCR has undergone tremendous improvements, and many variations on the basic PCR theme have been published. With such a large volume of PCR-related literature, a clinical scientist wishing to use the technique will have a difficult task loc- ing the relevant information to implement it effectively. There is thus clearly a need for an up-to-date volume with detailed protocols to facilitate the setting up of those techniques most relevant to clinical applications. Unlike some other books on this topic, Clinical Applications of PCR includes only methods that are of direct relevance in clinical settings. The book is organized in three parts: an introductory section, a section on general methodology, and a final section with specific clinical applications. The first section covers the basic principles of PCR and is most useful to those new to molecular diagnosis. The next chapter includes useful tips for setting up a PCR laboratory. Section 2 then outlines some of the most commonly used PCR-based techniques in molecular diagnosis. Section 3 includes carefully chosen examples that represent typical applications of PCR in diverse clinical fields, encompassing hematology, oncology, genetics, and microbiology.

Recenzje

". . . one of the first texts to carefully document all the needs required to perform PCR. . . [By] covering the fundamentals, methodologies, and applications of PCR, the editor has created a unique document that will be welcome in the clinical laboratory."-Doody's Health Sciences Book Review Journal

"Each chapter is clearly written and provides enough background material to explain the significance and application of the procedure. The protocols include sufficient detail to enable the reader to develop the test. This manual will be an important laboratory resource to those using PCR in the diagnosis of human diseases and should be of great interest to both molecular and clinical pathologists."-Archives of Pathology and Laboratory Medicine

Spis treści

Section 1: Introduction. Introduction to the Polymerase Chain Reaction, YMD LO. SETTING UP A PCR LABORATORY, YMD LO. Section 2: General Methodology. Amplification from Archival Materials, YMD Lo. Quantitative PCR, L Raemaekers. Mutation Screening Using PCR-SSCP: Silver Staining and Isotopic Protocols, PJ Saker. Multiple Fluorescence-Based PCR-SSCP Analysis with Primer-, Post-, and Internal Labeling, Hiroyuki Iwahana and Mitsuo Itakura. The Amplification Refractory Mutation System, YMD Lo. Artificial Restriction Fragment Length Polymorphism (A-RFLP) Analysis, YMD Lo and VA Horton. Long Range PCR, W Waggott. Generation of Labeled Probes by PCR, YMD Lo and SF An. PCR fo rthe Detection of Minority DNA Populations, YMD Lo. In Situ Amplification, John O' Leary. Sequencing of PCR Products, P Moss and SL Thein. Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR), R Kusec. Section 3: Clinical Applications. PCR Amplification of Minisatellite DNA for the Detection of Mixed Chimerism After Bone Marrow Transplantation, E Roux. Diagnosis of Mitochondrial Disorders Using the PCR, KJ Morten. PCR Analysis of CD-44 Variants in Tumors, J Bolodeoku. Detection of Circulating Solid Tumor Cells by Reverse Transcriptase Polymerase Chain Reaction, SA Burchill. Detection of Circulating Cancer Cells Using Immunobead-PCR, JE Hardingham. Single Cell PCR: Theory, Practice, and Clinical Applications, I Findlay. PCR-Based Noninvasive Prenatal Diagnosis Using Fetal Cells in Maternal Circulation, YMD Lo. Noninvasive Prenatal Diagnosis Using a Single Fetal Nucleated Erythrocyte Isolated by Micromanipulation from Maternal Blood, A Sekizawa, H Saito, and T Yanaihara. Diagnosis of Chromosomal Anueploidies Using Quantitative Fluorescent PCR, B Pertl and M Adinolfi. Detection of the Hepatitis C Virus by RT-PCR, CJ Healy and S Read. The Application of PCR to the Detection of M. tuberculosis in Sputum Samples, M Maher, M Glennon, M Cormican, and T Smith. Microsatellite Analysis in Human Disease, J Koreth, CJ Bakkenist, and J O' D. McGee. Index.

Blurb

Clinical Applications of PCR offers an unprecedented collection of core PCR techniques for the study and diagnosis of human diseases. Cutting-edge and essential for today's diagnostic laboratories, these techniques heavily utilize nonisotopic, solution phase, and in situ amplification methods. A significant number of chapters describe applications exploiting the exquisite sensitivity of PCR in the detection of rare or single cells, as in identifying fetal cells circulating in maternal blood, preimplantation embryo diagnosis, or detecting circulating cancer cells.
The methods described in Clinical Applications of PCR will well serve diverse clinical specialties ranging from hematology/oncology, human genetics, and microbiology, to virology, pathology, and infectious diseases. The book repeatedly demonstrates the power of PCR-its high sensitivity, specificity, and ability to rapidly discriminate sequence variations.