Clinical Applications of PCR

1. Front Matter

   Pages i-x

   PDF

2. Introduction

   1. Introduction to the Polymerase Chain Reaction

      • Y. M. Dennis Lo

      Pages 3-10

   2. Setting Up a PCR Laboratory

      • Y. M. Dennis Lo

      Pages 11-17

3. General Methodology

   1. Amplification from Archival Materials

      • Y. M. Dennis Lo

      Pages 21-25

   2. Quantitative PCR

      • Luc Raeymaekers

      Pages 27-38

   3. Mutation Screening Using PCR-SSCP

      • Philip J. Saker

      Pages 39-49

   4. Multiple Fluorescence-Based PCR-SSCP Analysis with Primer-, Post-, and Internal-Labeling

      • Hiroyuki Iwahana, Mitsuo Itakura

      Pages 51-60

   5. The Amplification Refractory Mutation System

      • Y. M. Dennis Lo

      Pages 61-69

   6. Artificial Restriction Fragment Length Polymorphism (A-RFLP) Analysis

      • Y. M. Dennis Lo, Virginia A. Horton

      Pages 71-79

   7. Long Range PCR

      • William Waggott

      Pages 81-91

   8. Generation of Labeled Probes by PCR

      • Y. M. Dennis Lo, Shu F. An

      Pages 93-100

   9. PCR for the Detection of Minority DNA Populations

      • Y. M. Dennis Lo

      Pages 101-107

   10. In Situ Amplification

       • John O’Leary

       Pages 109-144

   11. Sequencing of PCR Products

       • Paul Moss, Swee Lay Thein

       Pages 145-148

   12. Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR)

       • Rajko Kusec

       Pages 149-158

4. Clinical Application

   1. PCR Amplification of Minisatellite DNA for the Detection of Mixed Chimerism After Bone Marrow Transplantation

      • Etienne Roux

      Pages 161-170

   2. Diagnosis of Mitochondrial Disorders Using the PCR

      • Karl J. Morten

      Pages 171-187

   3. PCR Analysis of CD44 Variants in Tumors

      • John Bolodeoku

      Pages 189-202

   4. Detection of Circulating Solid Tumor Cells by Reverse Transcriptase Polymerase Chain Reaction

      • Susan A. Burchill

      Pages 203-224

   5. Detection of Circulating Tumor Cells Using lmmunobead-PCR

      • Jennifer E. Hardingham

      Pages 225-232

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.

". . . 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

• The Chinese University of Hong Kong, Hong Kong SAR

  Y. M. Dennis Lo

Policies and ethics