Visual detection of multiple genetically modified organisms in a capillary array

Ning Shao; Jianwei Chen; Jiaying Hu; Rong Li; Dabing Zhang; Shujuan Guo; Junhou Hui; Peng Liu; Litao Yang; Sheng-ce Tao

doi:10.1039/C6LC01330A

Visual detection of multiple genetically modified organisms in a capillary array†

Ning Shao,

‡^acd Jianwei Chen,‡^acd Jiaying Hu,‡^b Rong Li,^b Dabing Zhang,^b Shujuan Guo,^acd Junhou Hui,^e Peng Liu,^e Litao Yang*^b and Sheng-ce Tao*^acd

Author affiliations

* Corresponding authors

^a Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai 200240, China
E-mail: taosc@sjtu.edu.cn

^b Collaborative Innovation center for biosafety of GMOs, National Center for the Molecular Characterization of Genetically Modified Organisms, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
E-mail: yylltt@sjtu.edu.cn

^c State Key Laboratory of Oncogenes and Related Genes, Shanghai 200240, China

^d School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China

^e Department of Biomedical Engineering, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, School of Medicine, Tsinghua University, Beijing 100084, China

Abstract

There is an urgent need for rapid, low-cost multiplex methodologies for the monitoring of genetically modified organisms (GMOs). Here, we report a [C with combining low line] apillary [A with combining low line] rray-based [L with combining low line] oop-mediated isothermal amplification for [M with combining low line] ultiplex visual detection of nucleic acids (CALM) platform for the simple and rapid monitoring of GMOs. In CALM, loop-mediated isothermal amplification (LAMP) primer sets are pre-fixed to the inner surface of capillaries. The surface of the capillary array is hydrophobic while the capillaries are hydrophilic, enabling the simultaneous loading and separation of the LAMP reaction mixtures into each capillary by capillary forces. LAMP reactions in the capillaries are then performed in parallel, and the results are visually detected by illumination with a hand-held UV device. Using CALM, we successfully detected seven frequently used transgenic genes/elements and five plant endogenous reference genes with high specificity and sensitivity. Moreover, we found that measurements of real-world blind samples by CALM are consistent with results obtained by independent real-time PCRs. Thus, with an ability to detect multiple nucleic acids in a single easy-to-operate test, we believe that CALM will become a widely applied technology in GMO monitoring.