Signal Amplification

Signal amplification methods have been available in serology for many years and are characterized by increases in the detection of a target molecule rather than the molecule itself. Similar principles are used when detecting various nucleic acid targets. Two approaches of signal amplification methods involve nonenzymatic or enzymatic technologies. In nonenzy-matic signal amplification, the target is immobilized or localized using complementary probes. Ultimately, the hybrid molecule is detected using molecules labeled with alkaline phosphatase that can generate a colorimetric or chemiluminescence reaction (Fig. 6A and 6B). Alternatively, fluorescent probes can be used to directly visualize the hybrid molecule for in situ applications (Fig. 7). Signal amplification techniques are not prone to the problems of amplicon contamination because the target is not amplified. The other category of signal amplification uses enzymatic activities to replicate or generate a reporter molecule. These enzymatic signal amplification methods can be susceptible to false-positive reactions if the reporter molecule inadvertently contaminates the specimen. As a result, utmost care must be exercised when disposing of the specimen following detection.

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