Microchimica Acta

, 186:243 | Cite as

Colorimetric detection of nucleic acid sequences in plant pathogens based on CRISPR/Cas9 triggered signal amplification

  • Weidan Chang
  • Weipeng Liu
  • Ying Liu
  • Fangfang Zhan
  • Huifang Chen
  • Hongtao Lei
  • Yingju LiuEmail author
Original Paper


A colorimetric method is presented for the detection of specific nucleotide sequences in plant pathogens. It is based on the use of CRISPR/Cas9-triggered isothermal amplification and gold nanoparticles (AuNPs) as optical probes. The target DNA was recognized and broken up by a given Cas9/sgRNA complex. After isothermal amplification, the product was hybridized with oligonucleotide-functionalized AuNPs. This resulted in the aggregation of AuNPs and a color change from wine red to purple. The visual detection limit is 2 pM of DNA, while a linear relationship exists between the ratio of absorbance at 650 and 525 nm and the DNA concentration in the range from 0.2 pM to 20 nM. In contrast to the previous CRISPR-based amplification platforms, the method has significantly higher specificity with the single-base mismatch and can be visually read out. It was successfully applied to identify the Phytophthora infestans genomic DNA.

Graphical abstract

Schematic presentation of a colorimetric method for detection of Phytophthora infestans genomic DNA based on CRISPR/Cas9-triggered isothermal amplification. The Cas9 endonuclease cleaves DNA at the design site and the color changes from red to purple with increasing target DNA concentration.


Isothermal amplification Gold nanoparticles AuNP probes Triggered aggregation Single-base mismatch Phytophthora infestans Cas9/sgRNA complex Localized surface plasmon resonance Rolling circle amplification Double-strand break 



This work was supported by the National Scientific Foundation of China (21705051, 21874048), the Scientific Foundation of Guangdong Province (2017A030313077), the Science and Technology Planning Project of Guangdong Province (2016B030303010), National Key Research and Development Program of China (SQ2017YFC160089), the Program for the Top Young Innovative Talents of Guangdong Province (2016TQ03N305), and the Foundation for High-level Talents in South China Agricultural University.

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2019_3348_MOESM1_ESM.docx (987 kb)
ESM 1 (DOCX 986 kb)


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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Weidan Chang
    • 1
    • 2
  • Weipeng Liu
    • 1
  • Ying Liu
    • 1
  • Fangfang Zhan
    • 3
  • Huifang Chen
    • 1
  • Hongtao Lei
    • 2
  • Yingju Liu
    • 1
    Email author
  1. 1.College of Materials & EnergySouth China Agricultural UniversityGuangzhouChina
  2. 2.The Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food ScienceSouth China Agricultural UniversityGuangzhouChina
  3. 3.Fujian Key Laboratory of Plant Virology, Institute of Plant VirologyFujian Agriculture and Forestry UniversityFuzhouChina

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