Performance of probe polymerization-conjunction-agarose gel electrophoresis in the rapid detection of KRAS gene mutation
Abstract 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 thro...
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| Format: | Article |
| Language: | English |
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Sociedade Brasileira de Genética
2018-07-01
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| Series: | Genetics and Molecular Biology |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572018000400555&lng=en&tlng=en |
| _version_ | 1818215429573705728 |
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| author | Na Xiao Yi-Tong Tang Zhi-Shan Li Rui Cao Rong Wang Jiu-Ming Zou Jiao Pei |
| author_facet | Na Xiao Yi-Tong Tang Zhi-Shan Li Rui Cao Rong Wang Jiu-Ming Zou Jiao Pei |
| author_sort | Na Xiao |
| collection | DOAJ |
| description | Abstract 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 sequencing 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. |
| first_indexed | 2024-12-12T06:35:56Z |
| format | Article |
| id | doaj.art-ee7490b538d84e90a79f63a2470f841f |
| institution | Directory Open Access Journal |
| issn | 1678-4685 |
| language | English |
| last_indexed | 2024-12-12T06:35:56Z |
| publishDate | 2018-07-01 |
| publisher | Sociedade Brasileira de Genética |
| record_format | Article |
| series | Genetics and Molecular Biology |
| spelling | doaj.art-ee7490b538d84e90a79f63a2470f841f2022-12-22T00:34:28ZengSociedade Brasileira de GenéticaGenetics and Molecular Biology1678-46852018-07-0141355556110.1590/1678-4685-gmb-2017-0197S1415-47572018000400555Performance of probe polymerization-conjunction-agarose gel electrophoresis in the rapid detection of KRAS gene mutationNa XiaoYi-Tong TangZhi-Shan LiRui CaoRong WangJiu-Ming ZouJiao PeiAbstract 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 sequencing 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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572018000400555&lng=en&tlng=enMutationcirculating DNApolymerization-conjunction reactionagarose gel electrophoresisK-ras |
| spellingShingle | Na Xiao Yi-Tong Tang Zhi-Shan Li Rui Cao Rong Wang Jiu-Ming Zou Jiao Pei Performance of probe polymerization-conjunction-agarose gel electrophoresis in the rapid detection of KRAS gene mutation Genetics and Molecular Biology Mutation circulating DNA polymerization-conjunction reaction agarose gel electrophoresis K-ras |
| title | Performance of probe polymerization-conjunction-agarose gel electrophoresis in the rapid detection of KRAS gene mutation |
| title_full | Performance of probe polymerization-conjunction-agarose gel electrophoresis in the rapid detection of KRAS gene mutation |
| title_fullStr | Performance of probe polymerization-conjunction-agarose gel electrophoresis in the rapid detection of KRAS gene mutation |
| title_full_unstemmed | Performance of probe polymerization-conjunction-agarose gel electrophoresis in the rapid detection of KRAS gene mutation |
| title_short | Performance of probe polymerization-conjunction-agarose gel electrophoresis in the rapid detection of KRAS gene mutation |
| title_sort | performance of probe polymerization conjunction agarose gel electrophoresis in the rapid detection of kras gene mutation |
| topic | Mutation circulating DNA polymerization-conjunction reaction agarose gel electrophoresis K-ras |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572018000400555&lng=en&tlng=en |
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