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1.
Chembiochem ; 22(2): 392-397, 2021 01 15.
Artículo en Inglés | MEDLINE | ID: mdl-32881235

RESUMEN

With the advent of innovative genomic discovery toolkits such as RT-PCR, genetic information can be quickly decrypted, and this has resulted in significant progress in overcoming diseases. However, RT-PCR has the serious problem of frequent errors, and the demand for a new gene diagnostic system is emerging. Herein, we propose a universal coding system for the effective detection of short single-stranded DNA or RNA by using a topological transformation-based nano-barcoding technique (TNT). Our goal was to develop a dedicated diagnostic device that unifies the other gene groups, thus resulting in minimum testing. In a universal coding system consisting of two separate circulation structures, different gene groups become generalized into specific single genes with the same sequence by a strand-displacement reaction and are then amplified, eventually being quickly detected in one TNT system. Simple gene diagnostic systems like this make high-speed, point-of-care diagnostic technologies, and we are very confident that these will provide clinical gene detection in the near future.


Asunto(s)
Pruebas en el Punto de Atención , ARN/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Humanos
2.
Chembiochem ; 21(17): 2533-2539, 2020 09 01.
Artículo en Inglés | MEDLINE | ID: mdl-32291863

RESUMEN

As the market for personalized lung cancer medicine expands, the demand for molecular diagnostic tools in general, and methods of detecting multiple genes with qualitative, quantitative, and high specificity in particular, have grown. Here, we propose a system for the effective detection of lung cancer-specific, long-length epidermal growth factor receptor (EGFR) gene mutations by using a topological transformation nano-barcoding technique (TNT). In former TNT studies, EGFR was successfully detected in cell environments and at test stages in the presence of a reference gene. However, because typical EGFR target concentrations are significantly lower at the clinical stage and the probe-binding ability of long-length targets is lower that of short targets, our system employs polymerase chain reaction (PCR) amplification, restriction, and filtering (PRF) for EGFR fragmentation to maximize performance. In a PRF system, the target is amplified by PCR, cut to a suitable size by a restriction enzyme, and filtered by a magnetic bead. With detection limits of 0.3555 % and 1.500 % for EGFR Del 19 and L858R mutations, respectively, the proposed TNT with PRF can effectively distinguish mutant cell lines and efficiently detect various lengths of genetic variations in clinical trials.


Asunto(s)
Código de Barras del ADN Taxonómico , Receptores ErbB/genética , Grafito/química , Reacción en Cadena de la Polimerasa , Animales , Células Cultivadas , Grafito/síntesis química , Humanos , Ratones
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