A Molecular Review of the Detection of Specific Nucleic Acids by Amplification and Hybridization Characterization of Microbial Diversity in the Food Chain: A Molecular Review

Abstract

The ideal target DNA or RNA region used in nucleic acid detection in food safety should be appropriate in length, suitable for the design of primers and probes, and include minimal repeat sequences. However, in some cases, specific nucleic acid targets as short as approximately 20 nt or consisting of short tandem repeats must be determined, with microRNA and telomeres as two actual examples. In this chapter, specific nucleic acid detection methods involving amplification and hybridization will be reviewed. For the detection of miRNA, microarray chip methods, Northern blot methods, real-time PCR-based methods, in situ hybridization-based methods, and biosensor-based methods have been developed to overcome the too short length of miRNA. For the detection of telomere length, terminal restriction fragment analysis, quantitative PCR-based telomere detection, single telomere length analysis (STELA), and fluorescence in situ hybridization-based methods have been used. The advantages and disadvantages of these methods are analyzed in this review.