MSN/NA-doped nanoflower enhancing isothermal fluorescent sensor with a portable PCR tube fluorescence reader for the on-site detection of Vibrio parahaemolyticus

Yuan Su; Xuetong Li; Longjiao Zhu; Huashuo Chu; Yangzi Zhang; Jingjing Tian; Shuxia Lyu; Wentao Xu

doi:10.1016/j.aca.2022.339448

MSN/NA-doped nanoflower enhancing isothermal fluorescent sensor with a portable PCR tube fluorescence reader for the on-site detection of Vibrio parahaemolyticus

Anal Chim Acta. 2022 Apr 1:1200:339448. doi: 10.1016/j.aca.2022.339448. Epub 2022 Jan 6.

Authors

Yuan Su¹, Xuetong Li², Longjiao Zhu¹, Huashuo Chu¹, Yangzi Zhang¹, Jingjing Tian¹, Shuxia Lyu³, Wentao Xu⁴

Affiliations

¹ Key Laboratory of Precision Nutrition and Food Quality, Beijing Laboratory for Food Quality and Safety, Department of Nutrition and Health, China Agricultural University, Beijing 100191, China.
² College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang 110866, China.
³ College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang 110866, China. Electronic address: lushuxia@syau.edu.cn.
⁴ Key Laboratory of Precision Nutrition and Food Quality, Beijing Laboratory for Food Quality and Safety, Department of Nutrition and Health, China Agricultural University, Beijing 100191, China. Electronic address: xuwentao@cau.edu.cn.

PMID: 35256145
DOI: 10.1016/j.aca.2022.339448

Abstract

To develop an on-site, thermostatic and rapid sensor for the detection of Vibrio parahaemolyticus (V. parahaemolyticus), A single cross-priming fluorescence (SCPF) sensor was designed using a 3D nano-nucleic acid hybrid material that termed mesoporous silica nanoparticle/nucleic acid-doped nanoflower (MSN/NA-doped nanoflower). In addition, a portable polymerase chain reaction (PCR) tube fluorescence reader was built. Further analysis of the MSN/NA-doped nanoflower morphology and the enhancement mechanism indicated that the MSNs aggregated into larger nanoclusters by adsorbing single cross-priming amplification (sCPA) components, forming MSNs/NAs-doped nanoflowers, and increasing the local concentrations to enhance sCPA efficiency. Cyclic amplification relied mainly on the self-folding hairpin-like structure of the amplified product that continuously formed, opened, re-formed, and opened again. The target DNA was detected with a detection limit of 2.4 copies/μL.

Keywords: Cross-priming; Fluorescence reader; MSNs; Nanoflower; On-site; PCR tube; Vibrio parahaemolyticus.

MeSH terms

Nucleic Acid Amplification Techniques
Polymerase Chain Reaction
Sensitivity and Specificity
Vibrio parahaemolyticus* / genetics