Integrated microneedle-smartphone nucleic acid amplification platform for in-field diagnosis of plant diseases

Rajesh Paul; Emily Ostermann; Yuting Chen; Amanda C Saville; Yuming Yang; Zhen Gu; Anna E Whitfield; Jean B Ristaino; Qingshan Wei

doi:10.1016/j.bios.2021.113312

Integrated microneedle-smartphone nucleic acid amplification platform for in-field diagnosis of plant diseases

Biosens Bioelectron. 2021 Sep 1:187:113312. doi: 10.1016/j.bios.2021.113312. Epub 2021 May 8.

Authors

Rajesh Paul¹, Emily Ostermann¹, Yuting Chen², Amanda C Saville², Yuming Yang³, Zhen Gu⁴, Anna E Whitfield⁵, Jean B Ristaino⁵, Qingshan Wei⁶

Affiliations

¹ Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, 27696, USA.
² Department of Entomology and Plant Pathology, North Carolina State University, Raleigh, NC, 27695, USA.
³ Department of Agrotechnology and Food Sciences, Wageningen University, 6708, PB, Wageningen, Netherlands.
⁴ College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, China; Zhejiang Laboratory of Systems and Precision Medicine, Zhejiang University Medical Cencter, Hangzhou, Zhejing, 310058, China; Deparment of General Surgery, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, 310058, China.
⁵ Department of Entomology and Plant Pathology, North Carolina State University, Raleigh, NC, 27695, USA; Emerging Plant Disease and Global Food Security Cluster, North Carolina State University, Raleigh, NC, 27696, USA.
⁶ Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, 27696, USA; Emerging Plant Disease and Global Food Security Cluster, North Carolina State University, Raleigh, NC, 27696, USA. Electronic address: qwei3@ncsu.edu.

PMID: 34004545
DOI: 10.1016/j.bios.2021.113312

Abstract

We demonstrate an integrated microneedle (MN)-smartphone nucleic acid amplification platform for "sample-to-answer" diagnosis of multiplexed plant pathogens within 30 min. This portable system consists of a polymeric MN patch for rapid nucleic acid extraction within a minute and a 3D-printed smartphone imaging device for loop-mediated isothermal amplification (LAMP) reaction and detection. We expanded the extraction of the MN technology for DNA targets as in the previous study (ACS Nano, 2019, 13, 6540-6549) to more fragile RNA biomarkers, evaluated the storability of the extracted nucleic acid samples on MN surfaces, and developed a smartphone-based LAMP amplification and fluorescent reader device that can quantify four LAMP reactions on the same chip. In addition, we have found that the MN patch containing as few as a single needle tip successfully extracted enough RNA for RT-PCR or RT-LAMP analysis. Moreover, MN-extracted RNA samples remained stable on MN surfaces for up to three days. The MN-smartphone platform has been used to detect both Phytophthora infestans DNA and tomato spotted wilt virus (TSWV) RNA down to 1 pg, comparable to the results from a benchtop thermal cycler. Finally, multiplexed detection of P. infestans and TSWV through a single extraction from infected tomato leaves and amplification on the smartphone without benchtop equipment was demonstrated.

Keywords: Isothermal amplification; Microneedle; Nucleic acid extraction; Plant disease diagnostics; Smartphone.

MeSH terms

Biosensing Techniques*
Molecular Diagnostic Techniques
Nucleic Acid Amplification Techniques
Plant Diseases
Smartphone*

Supplementary concepts

LAMP assay