Single nucleotide polymorphism (SNP) refers to the polymorphism of DNA sequence caused by the conversion, transposition, insertion or deletion of single nucleotide at the genomic level, which is directly related to many diseases. Therefore, the development of fast, accurate, and inexpensive SNP detection technology is essential for drug research, personalized medicine, clinical trials, and molecular diagnosis. Most current SNP detection methods require complex probe labeling and rely on High-cost testing equipment. Tang Zhuo's research group of Chengdu Institute of Biology, Chinese Academy of Sciences, has taken organic chemistry, nucleic acid chemistry and chemical biology as research directions, and has been devoted to the development of nucleic acid detection biosensors based on ribozymes. Recently, a new Gap-LCR's SNP detection technology.
Tang Zhuo's group introduced a short functional DNA sequence (DNAzyme) to the Gap-LCR probe, thereby avoiding chemical modification of the probe; then using two exonucleases to sequentially degrade the system without ligation reaction Phosphorylation probe and release the DNAzyme sequence in the Gap-LCR amplification product; finally, the combination of DNAzyme and porphyrin iron (hemin) or porphyrin iron analogue has properties similar to peroxidase activity, catalyzing H202 oxidation Substrates (ABTS, tyramine hydrochloride, etc.) become colored or fluorescent substances that can be detected directly at room temperature through changes in color or fluorescent signal. In the end, the researchers obtained a high-sensitivity and specific SNP detection technology with a detection limit of 1,000 molecules and a target molecule from 100,000 interfering molecules.
This study verified the sensitivity, specificity and versatility of the method. The researchers successfully applied the detection technique to the detection of the most common mutation site GJB2 235delC site in neonatal deafness genes in blood DNA samples.
Related research results were published in the journals Biosensors and Bioelectronics.
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