15 – A Chemiluminescent DNA Probe Test Based on Strand Displacement Amplification

This chapter introduces a DNA probe test that is based on strand displacement amplification (SDA) and chemiluminescence detection. A step-by-step protocol is presented for amplification and detection of a DNA sequence specific to Myobacterium tuberculosis, the causative agent of tuberculosis. The chapter also covers application of the protocol to clinical sputum specimens. The Myobacterium tuberculosis system provides a model for development of SDA and chemiluminescence detection systems for viruses and other infectious agents. SDA requires sequence-specific detection methods. Analysis of SDA products using agarose gel electrophoresis and ethidium staining, a technique commonly used for polymerase chain reaction, routinely reveals a number of non-target amplification products that obscure identification of target amplification. Application of this system to other targets require modifications of the sample processing protocol, the DNA sequences, the reaction conditions of SDA, and the detection assay.