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Competitive activation cross amplification combined with smartphone-based quantification for point-of-care detection of single nucleotide polymorphism

文献类型： 外文期刊

作者： Wen, Junping 1 ; Gou, Hongchao 2 ; Wang, Siyuan 3 ; Lin, Qijie 1 ; Chen, Kaifeng 1 ; Wu, Yuqian 1 ; Huang, Xuehuan 1 ; Sh 1 ;

作者机构： 1.South China Agr Univ, Natl & Reg Joint Engn Lab Medicament Zoonoses Pre, Key Lab Zoonoses,Key Lab Anim Vaccine Dev,Guangdo, Minist Agr,Key Lab Zoonoses Prevent & Control Gua, Guangzhou 510642, Peoples R China

2.Guangdong Acad Agr Sci, Inst Anim Hlth, Guangzhou 510640, Peoples R China

3.China Agr Univ, Minist Agr, Key Lab Agr Informat Acquisit Technol, Beijing 100083, Peoples R China

4.China Agr Univ, Minist Educ, Key Lab Modern Precis Agr Syst Integrat Res, Beijing 100083, Peoples R China

关键词： Single nucleotide polymorphism; Competitive activation cross amplification; RNase H2 enzyme; Smartphone; Point-of-care testing

期刊名称：BIOSENSORS & BIOELECTRONICS （ 影响因子：10.257； 五年影响因子：8.669 ）

ISSN： 0956-5663

年卷期： 2021 年 183 卷

页码：

收录情况： SCI

摘要： In this study, we firstly propose a novel smartphone-assisted visualization SNP genotyping method termed competitive activation cross amplification (CACA). The mutation detection strategy depends on the ingenious design of both a start primer and a verification probe with ribonucleotide insertion through competitive combination and perfect matching with the target DNA, Meanwhile, the RNase H2 enzyme was utilized to specifically cleave ribonucleotide insertion and achieve extremely specific dual verification. Simultaneously, the results allow both colorimetric and fluorescence product dual-mode visualization by using self-designed 3D-printed dual function cassette. We validated this novel CACA by analyzing the Salmonella Pullorum rfbS gene at the 237th site, successfully solve the current bottleneck of specific identification and visual detection of this pathogen. The concentration detection limits of the plasmid and genomic DNA were 1500 copies/?L and 3.98 pg/?L, respectively, and as low as the presence of 0.1% mutant-type can be distinguished from 99.9% wild-type. Combined with a powerful hand-warmer, which can provide heating more than 60 ?C for 20 h to realize power-free, dual function cassette and smartphone quantitation, our novel CACA platform firstly realizes user-friendly, costeffective, portable, rapid, and accurate POC detection of SNP.

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