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.
Inhaltsverzeichnis
to the Polymerase Chain Reaction. - Setting Up a PCR Laboratory. - General Methodology. - Amplification from Archival Materials. - Quantitative PCR. - Mutation Screening Using PCR-SSCP. - Multiple Fluorescence-Based PCR-SSCP Analysis with Primer-, Post-, and Internal-Labeling. - The Amplification Refractory Mutation System. - Artificial Restriction Fragment Length Polymorphism (A-RFLP) Analysis. - Long Range PCR. - Generation of Labeled Probes by PCR. - PCR for the Detection of Minority DNA Populations. - In Situ Amplification. - Sequencing of PCR Products. - Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). - Clinical Application. - PCR Amplification of Minisatellite DNA for the Detection of Mixed Chimerism After Bone Marrow Transplantation. - Diagnosis of Mitochondrial Disorders Using the PCR. - PCR Analysis of CD44 Variants in Tumors. - Detection of Circulating Solid Tumor Cells by Reverse Transcriptase Polymerase Chain Reaction. - Detection of Circulating Tumor Cells Using lmmunobead-PCR. - Single Cell PCR. - PCR-Based Noninvasive Prenatal Diagnosis Using Fetal Cells in Maternal Circulation. - Noninvasive Prenatal Diagnosis Using a Single Fetal Nucleated Erythrocyte Isolated by Micromanipulation from Maternal Blood. - Diagnosis of Chromosomal Aneuploidies Using Quantitative Fluorescent PCR. - Detection of the Hepatitis C Virus by RT-PCR. - The Application of PCR to the Detection of M. tuberculosis in Sputum Samples. - Microsatellite Analysis in Human Disease.
