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The genetic authentication of Panax ginseng and Panax quinquefolius based on using single nucleotide polymorphism (SNP) conducted in a nucleic acid test chip

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Abstract

Panax ginseng and Panax quinquefolius, which are commonly called Chinese ginseng and American ginseng respectively, have different medicinal properties and market values; however, these samples can be difficult to differentiate from one another based on physical appearances of the samples especially when they are in powdery or granular forms. A molecular technique is thus needed to overcome this difficulty; this technique is based on the nucleic acid test (NAT) conducted on the microfluidic chip surface. Three single nucleotide polymorphism (SNP) sites (i.e. N1, N2, N3) on the Panax genome that differ between P. ginseng (G) and P. quinquefolius (Q) have been selected to design probes for the NAT. Primers were designed to amplify the antisense strands by asymmetric PCR. We have developed three different NAT methodologies involving surface immobilization and subsequent (stop flow or dynamic) hybridization of probes (i.e. N1G, N1Q, N2G, N2Q, N3Q) to the antisense strands. These NAT methods consist of two steps, namely immobilization and hybridization, and each method is distinguished by what is immobilized on the microfluidic chip surface in the first step (i.e. probe, target or capture strand). These three NATs developed are called probe-target method 1, target-probe method 2 and three-strand complex method 3. Out of the three methods, it was found that the capture strand-target-probe method 3 provided the best differentiation of the ginseng species, in which a 3' NH2 capture strand is first immobilized and the antisense PCR strand is then bound, while N2G and N3Q probes are used for detection of P. ginseng (G) and P. quinquefolius (Q) respectively.

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Funding

This work received funding from NSERC and Macan Biotechnologies Ltd. of Macau.

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Author notes

  1. Abootaleb Sedighi

    Present address: Department of Chemistry, University of Toronto, Toronto, ON, Canada

Authors and Affiliations

  1. Department of Chemistry, Simon Fraser University, 8888 University Drive, Burnaby, BC, V5A 1S6, Canada

    Christopher Oberc, Abootaleb Sedighi & Paul C. H. Li

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  1. Christopher Oberc
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  2. Abootaleb Sedighi
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Contributions

Christopher Oberc contributed by designing and conducting all experiments and wrote the paper; Abootaleb Sedighi provided advice on designing experiments; Paul C.H. Li, designed the experiments, wrote and revised the paper.

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Correspondence to Paul C. H. Li.

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Oberc, C., Sedighi, A. & Li, P.C.H. The genetic authentication of Panax ginseng and Panax quinquefolius based on using single nucleotide polymorphism (SNP) conducted in a nucleic acid test chip. Anal Bioanal Chem 414, 3987–3998 (2022). https://doi.org/10.1007/s00216-022-04044-0

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  • DOI: https://doi.org/10.1007/s00216-022-04044-0

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