DNA Authentication of St John’s Wort (Hypericum perforatum L.) Commercial Products Targeting the ITS Region
| Autor: | Tiziana Sgamma, Purvi Mali, Adrian Slater, Eva Masiero, Marco Kreuzer, Eleanor Hill, Caroline Howard |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Hypericum perforatum lcsh:QH426-470 DNA fragmentation Biology 01 natural sciences DNA sequencing 03 medical and health sciences chemistry.chemical_compound Genetics DNA barcoding Internal transcribed spacer Fragmentation (cell biology) Genetics (clinical) St John’s Wort Ribosomal RNA barcoding 0104 chemical sciences 010404 medicinal & biomolecular chemistry qPCR lcsh:Genetics 030104 developmental biology Biochemistry chemistry NGS metabarcoding Primer (molecular biology) medicinal plant extract DNA |
| Zdroj: | Genes, Vol 10, Iss 4, p 286 (2019) Genes Volume 10 Issue 4 |
| ISSN: | 2073-4425 |
| Popis: | There is considerable potential for the use of DNA barcoding methods to authenticate raw medicinal plant materials, but their application to testing commercial products has been controversial. A simple PCR test targeting species-specific sequences within the nuclear ribosomal internal transcribed spacer (ITS) region was adapted to screen commercial products for the presence of Hypericum perforatum L. material. DNA differing widely in amount and extent of fragmentation was detected in a number of product types. Two assays were designed to further analyse this DNA using a curated database of selected Hypericum ITS sequences: A qPCR assay based on a species-specific primer pair spanning the ITS1 and ITS2 regions, using synthetic DNA reference standards for DNA quantitation and a Next Generation Sequencing (NGS) assay separately targeting the ITS1 and ITS2 regions. The ability of the assays to detect H. perforatum DNA sequences in processed medicines was investigated. Out of twenty different matrices tested, both assays detected H. perforatum DNA in five samples with more than 103 ITS copies µ L&minus 1 DNA extract, whilst the qPCR assay was also able to detect lower levels of DNA in two further samples. The NGS assay confirmed that H. perforatum was the major species in all five positive samples, though trace contaminants were also detected. |
| Databáze: | OpenAIRE |
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