A signal-flexible gene diagnostic strategy coupling loop-mediated isothermal amplification with hybridization chain reaction

Qing Dong; Quanyi Liu; Lulu Guo; Dan Li; Xudong Shang; Bingling Li; Yan Du

doi:10.1016/j.aca.2019.06.048

A signal-flexible gene diagnostic strategy coupling loop-mediated isothermal amplification with hybridization chain reaction

Anal Chim Acta. 2019 Nov 4:1079:171-179. doi: 10.1016/j.aca.2019.06.048. Epub 2019 Jun 29.

Authors

Qing Dong¹, Quanyi Liu², Lulu Guo², Dan Li³, Xudong Shang², Bingling Li⁴, Yan Du⁵

Affiliations

¹ State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022, China; Changchun University of Science and Technology, Changchun, Jilin, 130022, China.
² State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022, China; Department of Chemistry, University of Science & Technology of China, Hefei, Anhui, 230026, China.
³ State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022, China. Electronic address: lidan@ciac.ac.cn.
⁴ State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022, China; Department of Chemistry, University of Science & Technology of China, Hefei, Anhui, 230026, China. Electronic address: binglingli@ciac.ac.cn.
⁵ State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022, China; Department of Chemistry, University of Science & Technology of China, Hefei, Anhui, 230026, China. Electronic address: duyan@ciac.ac.cn.

Abstract

Recent study proves that the combination of loop mediated isothermal nucleic acid amplification (LAMP) with one-step strand displacement (OSD) is of great help to improve the sequence specificity during genetic detection. However, because OSD is incapable of signal amplification, the signal-to-noise ratio or the observable signal change may be usually not significant enough to satisfy practical usage. With the purpose to overcome this challenge, herein a more advanced and practical sensing principle is developed with the OSD replaced by an amplifiable nucleic acid circuit, hybridization chain reaction (HCR). The very contagious norovirus (NoV) was employed as the model target. Compared with LAMP-OSD, the LAMP-HCR can detect as few as 30 copies of NoV gene in 2% fecal samples with significantly enlarged signal change and signal-to-background ratio. Therefore, more reliable detection is achieved. Moreover, due to the high compatibility of HCR, the final LAMP-HCR products can be flexibly transduced into different types of readouts, including fluorescence, flow cytometer (FCM) and even a personal glucose meter (PGM). This further enlarges the operating environments for the detection from hospital labs, bedsides, to potential off-the-shelf devices in local pharmacies. Especially when using FCM or PGM, with the assistance of magnetic beads (MBs), the detection shows even higher tolerance capability to complicated biological matrices.

Keywords: Flow cytometer; Hybridization chain reaction; Loop-mediated isothermal amplification; Norovirus detection; Personal glucometer.

MeSH terms

DNA, Viral / analysis*
DNA, Viral / genetics
Feces / virology
Flow Cytometry / methods*
Genes, Viral*
Humans
Magnetic Phenomena
Norovirus / genetics*
Nucleic Acid Amplification Techniques / methods
Nucleic Acid Hybridization
Reproducibility of Results

Substances

DNA, Viral