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Dark field microscope-based single nanoparticle identification coupled with statistical analysis for ultrasensitive biotoxin detection in complex sample matrix

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Abstract

A novel approach for ultrasensitive ochratoxin A (OTA) detection is reported based on dark field microscope-based single nanoparticle identification coupled with a statistical analysis method. OTA aptamers were firstly hybridized with a single-stranded DNA (DNA1) to form an identification probe (DNA1-Apt). The aptamers separate from DNA1 in the presence of OTA and are released from the identification probe. Then, another single-stranded DNA (DNA2) hybridizes with DNA1 and result in the aggregation of gold nanoparticles (AuNPs). Therefore, the presence of AuNP aggregates is the evidence of the presence of OTA, while AuNP aggregates can be easily identified together with the monomers under dark field microscopic inspection. On the other hand, by counting the aggregation rate (the number of AuNP aggregates versus the number of AuNP monomers) with a statistical analysis method, OTA could be quantitatively detected. The detection range for OTA was 0.1 pg/mL ~ 30 ng/mL and the limit of detection was 0.1 pg/mL. The proposed sensor has comparative detection performance to sensors utilizing a number of signal amplification procedures, with the additional advantages of simplicity and high efficiency. The sensor can also be adopted for other target detection simply by replacing the identification probes.

The schematic of the AuNP aggregation for OTA detection. The OTA aptamers were competitively banded by OTA and induced form AuNP aggregation after adding DNA2 and AuNPs2. Subsequently, AuNPs were detected under dark field microscope and statistical analysis.

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Funding

This work was supported by the National Natural Science Foundation of China (21675028), the key Research and Development Program of Zhejiang Province (2020C02022), Thirteenth Five-Year Plan” Marine Economy Innovation and Development Demonstration Project (Grant No. FZHJ19), Nature Sciences Funding of Fujian Province (2018 J01682, 2019 J01238), STS Key Project of Fujian Province (2017 T3007), the Cooperative Project of Production and Study in University of Fujian Province (2018Y4007), and the Program for New Century Excellent Talents in Fujian Province University.

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Authors and Affiliations

  1. Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, 350116, Fujian, China

    Shaohua Xu, Fang Luo, Bin Qiu & Zhenyu Lin

  2. College of Biological, Chemical Sciences and Engineering, Jiaxing University, Jiaxing, 314001, China

    Longhua Guo & Lifen Chen

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  1. Shaohua Xu
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Correspondence to Longhua Guo, Lifen Chen or Bin Qiu.

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Xu, S., Guo, L., Chen, L. et al. Dark field microscope-based single nanoparticle identification coupled with statistical analysis for ultrasensitive biotoxin detection in complex sample matrix. Microchim Acta 187, 413 (2020). https://doi.org/10.1007/s00604-020-04386-5

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