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L32 manuscript v19.pdf (546.66 kB)

Sensitive and Selective Detection of DNA Fragments Associated with Ganoderma Boninense by DNA-Nanoparticle Conjugate Hybridisation

preprint
revised on 14.07.2020 and posted on 14.07.2020 by Ekta Rani, Siti Akhtar Mohshim, Nor Hidayat Yusof, Muhammad Zamharir Ahmad, Royston Goodacre, Shahrul Ainliah Alang Ahmad, Lu Shin Wong

A colorimetric assay for the detection of DNA fragments associated with the oil palm pathogen Ganoderma boninense is reported, which is based on the aggregation of DNA-nanoparticle conjugated in the presence of complementary DNA from the pathogen. Here, various designs of DNA-nanoparticle conjugates were evaluated, and it was found that the best design gave a visually observable colour change with as little as 2 pmol of double-stranded DNA analyte even in the presence of a large excess of a mixture of non-complementary DNA. The assay was also able to differentiate analyte sequences with three or more single nucleotide mismatches. Overall, this label-free system is rapid, sensitive, selective, simple in design and easy to carry out. It does not require specialist equipment or specialist training for the interpretation of the results and therefore has the potential to be deployed of agricultural diagnostics in the field.

Funding

British Council 216196834

EPSRC EP/K024485/1

HEFCE N8 Agri-food Network

History

Email Address of Submitting Author

l.s.wong@manchester.ac.uk

Institution

University of Manchester

Country

United Kingdom

ORCID For Submitting Author

0000-0002-7437-123X

Declaration of Conflict of Interest

None

Version Notes

Latest version: 19

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