Dew inspired breathing-based detection of genetic point mutation visualized by naked eye

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Abstract

A novel label-free method based on breathing-induced vapor condensation was developed for detection of genetic point mutation. The dew-inspired detection was realized by integration of target-induced DNA ligation with rolling circle amplification (RCA). The vapor condensation induced by breathing transduced the RCA-amplified variances in DNA contents into visible contrast. The image could be recorded by a cell phone for further or even remote analysis. This green assay offers a naked-eye-reading method potentially applied for point-of-care liver cancer diagnosis in resource-limited regions.

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Most cited references 31

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Global cancer statistics

DM Parkin, P Pisani, J Ferlay … (2011)

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Mutation detection and single-molecule counting using isothermal rolling-circle amplification.

P M Lizardi, X. Huang, Z. Zhu … (1998)

Rolling-circle amplification (RCA) driven by DNA polymerase can replicate circularized oligonucleotide probes with either linear or geometric kinetics under isothermal conditions. In the presence of two primers, one hybridizing to the + strand, and the other, to the - strand of DNA, a complex pattern of DNA strand displacement ensues that generates 10(9) or more copies of each circle in 90 minutes, enabling detection of point mutations in human genomic DNA. Using a single primer, RCA generates hundreds of tandemly linked copies of a covalently closed circle in a few minutes. If matrix-associated, the DNA product remains bound at the site of synthesis, where it may be tagged, condensed and imaged as a point light source. Linear oligonucleotide probes bound covalently on a glass surface can generate RCA signals, the colour of which indicates the allele status of the target, depending on the outcome of specific, target-directed ligation events. As RCA permits millions of individual probe molecules to be counted and sorted using colour codes, it is particularly amenable for the analysis of rare somatic mutations. RCA also shows promise for the detection of padlock probes bound to single-copy genes in cytological preparations.

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Mutational hotspot in the p53 gene in human hepatocellular carcinomas.

I. C. Hsu, R Metcalf, T Sun … (1991)

Human hepatocellular carcinomas (HCC) from patients in Qidong, an area of high incidence in China, in which both hepatitis B virus and aflatoxin B1 are risk factors, were analysed for mutations in p53, a putative tumour-suppressor gene. Eight of the 16 HCC had a point mutation at the third base position of codon 249. The G----T transversion in seven HCC DNA samples and the G----C transversion in the other HCC are consistent with mutations caused by aflatoxin B1 in mutagenesis experiments. No mutations were found in exons 5,6,8 or the remainder of exon 7. These results contrast with p53 mutations previously reported in carcinomas and sarcomas of human lung, colon, oesophagus and breast; these are primarily scattered over four of the five evolutionarily conserved domains, which include codon 249 (refs 4-9). We suggest that the mutant p53 protein may be responsible for a selective clonal expansion of hepatocytes during carcinogenesis.

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Author and article information

Journal

Journal ID (nlm-ta): Sci Rep

Journal ID (iso-abbrev): Sci Rep

Title: Scientific Reports

Publisher: Nature Publishing Group

ISSN (Electronic): 2045-2322

Publication date (Electronic): 09 September 2014

Publication date Collection: 2014

Volume: 4

Electronic Location Identifier: 6300

Affiliations

[1 ]Department of Biomedical Engineering, School of Medicine, Tsinghua University , Beijing 100084, China

[2 ]Institute of Molecular Medicine, College of Life and Health Science, Northeastern University , Shenyang 110004, China

[3 ]Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases , Hangzhou 310003, China

[4 ]Tianjin Second People's Hospital and Tianjin Institute of Hepatology , Tianjin 300192, China

Author notes

[a ] duyanan@ 123456tsinghua.edu.cn

[b ] tshgl@ 123456tsinghua.edu.cn

Article

Publisher Item ID: srep06300

DOI: 10.1038/srep06300

PMC ID: 4158330

SO-VID: b214cdb8-df11-4b50-b8ce-27a615f924b7

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This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/

Dew inspired breathing-based detection of genetic point mutation visualized by naked eye

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Abstract

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Recursive Rule based Visual Categorization

Most cited references 31

Global cancer statistics

Mutation detection and single-molecule counting using isothermal rolling-circle amplification.

Mutational hotspot in the p53 gene in human hepatocellular carcinomas.

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