Nucleic Acid Hybridization and Amplification In Situ

Principles and Applications in Molecular Pathology
  • Matthew S. Cowlen
Part of the Pathology and Laboratory Medicine book series (PLM)


The ability of pathologists to diagnose disease has been enhanced significantly by the application of nucleic acid technologies, such as Southern blot hybridization and the polymerase chain reaction (PCR). These techniques are used routinely in clinical molecular diagnostic laboratories to detect and analyze genetic alterations associated with human disease. However, the ability to associate the nucleic acid sequence of interest with histopathologic or cytogenetic abnormalities directly is lost when tissue is destroyed during the extraction of nucleic acids. The analysis of nucleic acids in situ overcomes this limitation. ISH* offers the combined advantages of molecular biology, analytical morphology, and cytogenetics by facilitating the analysis of DNA or RNA in tissues and chromosomes. ISA of nucleic acid sequences using PCR is a developing technology that offers increased sensitivity compared to conventional ISH for detecting low-copy sequences. In situ nucleic acid techniques are discussed in detail in this chapter, with emphasis on the principles and clinical relevance of each technique.


Human Immunodeficiency Virus Type Chronic Lymphocytic Leukemia Comparative Genomic Hybridization Minimal Residual Disease Metaphase Chromosome 
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  • Matthew S. Cowlen

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