Performance of probe polymerization-conjunction-agarose gel electrophoresis in the rapid detection of KRAS gene mutation



Título del documento: Performance of probe polymerization-conjunction-agarose gel electrophoresis in the rapid detection of KRAS gene mutation
Revista: Genetics and molecular biology
Base de datos: PERIÓDICA
Número de sistema: 000419990
ISSN: 1415-4757
Autores: 1
1
2
3
2
2
2
Instituciones: 1Hubei University of Arts and Science, Medical College, Xianyang, Shaanxi. China
2Hubei University of Arts and Science, Department of Clinical Laboratory, Xianyang, Shaanxi. China
3Maternal and Child Health Care Hospital of Dongguan, Department of Internal Medicine, Dongguan. China
Año:
Periodo: Sep
Volumen: 41
Número: 3
Paginación: 555-561
País: Brasil
Idioma: Inglés
Tipo de documento: Artículo
Enfoque: Experimental
Resumen en inglés This study aimed to develop a simple and rapid method to detect KRAS gene mutations for conventional clinical applications under laboratory conditions. The genotype of mutation sites was determined based on the occurrence of target bands in the corresponding lanes of the reaction tubes through polymerization-conjunction of the probes, probe purification and amplification, and agarose gel electrophoresis. Circulating DNA samples were obtained from the plasma of 72 patients with lung cancer, which were identified based on six mutation sites (G12S, G12R, G12C, G12D, G12A, and G12V) of codon 12 of the KRAS gene. The detection results were compared with direct sequenc- ing data. The proposed detection method is characterized by simple operation, high specificity, and high sensitivity (2%). This method can detect the mutations of three samples at G12S, G12R, and G12A. In the direct sequencing spectra of these samples, the genotype could not be determined due to the lack of evident sequencing peaks that correspond to the basic group of mutations. In conclusion, a simple and rapid method was established based on probe polymerization-conjunction-agarose gel electrophoresis for detecting KRAS gene mutations. This method can be applied to the conventional mutation detection of inhomogeneous samples
Disciplinas: Medicina,
Química
Palabras clave: Genética,
Bioquímica,
Mutación,
ADN circulante,
Reacción de polimerización-conjunción,
Electroforesis,
Gel de agarosa,
Gen K-ras
Keyword: Genetics,
Biochemistry,
Mutation,
Circulating DNA,
Polymerization-conjunction reaction,
Electrophoresis,
Agarose gel,
K-ras gene
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