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Book Review |
ISBN 1-58829-549-4, Humana Press, Totowa, NJ; 2006; hardcover, pp 243, $99.50; also available in an e-book.
Molecular Genetic Technology Program,The University of Texas / M.D. Anderson Cancer Center, Houston, Texas
Address correspondence to Peter C. Hu, M.S., Molecular Genetic Technology Program, The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Blvd Unit 146, Houston, TX 77030, tel 713 563 3095; e-mail pchu{at}mdanderson.org.
Primed In Situ Hybridization (PRINS) was introduced about the same time as Fluorescent In Situ Hybridization (FISH). PRINS offers a faster detection method by making use of small unlabeled DNA sequences (oligonucleotides) that specifically bind to repeat sequences on the chromosomes (alpha satellite, Alu repeats, or telomeres) and then multiply exponentially through annealing and in situ elongation of the chromosomes. Visualization can take place by incorporating one labeled nucleotide into the newly generated fragments. Both of these techniques offer ways to detect chromosomal aneuploidies and subtle rearrangements. PRINS has been relatively neglected because FISH was easier to implement into the existing cytogenetic testing protocols. However, PRINS is making a come-back with modified techniques that offer faster detection, higher sensitivity, and greater specificity than FISH.
An interesting chapter concerns the ability to use PRINS to target specific sequences (Chapter 4). The target example used in this chapter is the SRY gene located on the Y chromosome for looking at women with XY gonadal dysgenesis and azospermic men with Xp-Yp interchange.This technique also has a potential role in non-invasive fetal gender screening. That is, instead of using amniocentesis where a large needle is inserted into the amnionic sac of the mother to withdraw fluid (which has circulating fetal cells) to test for fetal gender type, one can draw maternal blood (no threat to the fetus), which has circulating fetal cells, and apply a unique sequence PRINS procedure to find out the fetal gender. For small clinical labs or research labs that cannot afford a real-time instrument, this can serve as an alternative method that is equally effective, but less expensive,
Chapter 6 discusses using multi-color PRINS to look at human sperm for chromosome aneuploidy. is technique uses 5 fluorochromes to look at 3-4 different chromosomes simultaneously. In this technique each PRINS reaction of annealing and elongation lasts only 5 min. Currently, there is a FISH test (AneuVysion, made by Vysis) that looks at 5 of the most common aneuploidy chromosomes during pregnancy (ie, 13, 18, 21, X, and Y). These chromosomes account for a majority of miscarriages and multiple congenital malformation syndromes in liveborns. The FISH test, however, is performed on the growing fetus. Sperm aneuploidy PRINS tests spermatozoa for aneuploidy before conception even takes place. Other methods of studying spermatozoa have limitations in probe size and decondensation. The PRINS technique addresses these problems and appears to be more effective and specific than using FISH on sperm.
This book would be useful in an advanced genetics course. The discussions on theory and some of the chapters could readily be incorporated into existing advanced courses on cytogenetics or molecular techniques. Researchers will find this book useful since many techniques are introduced with specific protocols.The book has the potential to sprout new research ideas for student projects or larger scale projects. Some of the PRINS techniques may be applicable in the clinical setting for secondary validation tests with fast, specific, and highly sensitive detection methodologies.
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