title: Identification of Chinese Herbs Using a Sequencing-Free Nanostructured Electrochemical DNA Biosensor.
abstract: Due to the nearly identical phenotypes and chemical constituents, it is often very challenging to accurately differentiate diverse species of a Chinese herbal genus. Although technologies including DNA barcoding have been introduced to help address this problem, they are generally time-consuming and require expensive sequencing. Herein, we present a simple sequencing-free electrochemical biosensor, which enables easy differentiation between two closely related Fritillaria species. To improve its differentiation capability using trace amounts of DNA sample available from herbal extracts, a stepwise electrochemical deposition of reduced graphene oxide (RGO) and gold nanoparticles (AuNPs) was adopted to engineer a synergistic nanostructured sensing interface. By using such a nanofeatured electrochemical DNA (E-DNA) biosensor, two Chinese herbal species of Fritillaria (F. thunbergii and F. cirrhosa) were successfully discriminated at the DNA level, because a fragment of 16-mer sequence at the spacer region of the 5S-rRNA only exists in F. thunbergii. This E-DNA sensor was capable of identifying the target sequence in the range from 100 fM to 10 nM, and a detection limit as low as 11.7 fM (S/N = 3) was obtained. Importantly, this sensor was applied to detect the unique fragment of the PCR products amplified from F. thunbergii and F. cirrhosa, respectively. We anticipate that such a direct, sequencing-free sensing mode will ultimately pave the way towards a new generation of herb-identification strategies.
authors.affiliation: School of Laboratory Medicine, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, China. yanlei91316@163.com.|School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, China. yanlei91316@163.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: Lei
authors.affiliation: School of Laboratory Medicine, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, China. Yangf1013@163.com.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: School of Laboratory Medicine, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, China. hjtanglina@163.com.
authors.forename: Lina
authors.initials: L
authors.lastname: Tang
authors.affiliation: School of Pharmacy, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, China. kelichen@126.com.
authors.forename: Keli
authors.initials: K
authors.lastname: Chen
authors.affiliation: School of Laboratory Medicine, Hubei University of Chinese Medicine, 1 Huangjia Lake West Road, Wuhan 430065, China. zhanggj@hbtcm.edu.cn.
authors.forename: Guo-Jun
authors.initials: GJ
authors.lastname: Zhang
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D018744:DNA, Plant / Q000032:analysis* / Q000145:classification; D004365:Drugs, Chinese Herbal; D055664:Electrochemical Techniques / Q000379:methods*; D031410:Fritillaria / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry
chemical_list: D018744:DNA, Plant; D004365:Drugs, Chinese Herbal; D006046:Gold
keywords: Chinese herb; DNA identification; Fritillaria; electrochemical biosensor; nanomaterials-modified electrode