Rapid and sensitive identification of the herbal tea ingredient Taraxacum formosanum using loop-mediated isothermal amplification

Guan-Hua Lai; Jung Chao; Ming-Kuem Lin; Wen-Te Chang; Wen-Huang Peng; Fang-Chun Sun; Meng-Shiunn Lee; Meng-Shiou Lee

doi:10.3390/ijms16011562

Rapid and sensitive identification of the herbal tea ingredient Taraxacum formosanum using loop-mediated isothermal amplification

Int J Mol Sci. 2015 Jan 9;16(1):1562-75. doi: 10.3390/ijms16011562.

Authors

Guan-Hua Lai¹, Jung Chao², Ming-Kuem Lin³, Wen-Te Chang⁴, Wen-Huang Peng⁵, Fang-Chun Sun⁶, Meng-Shiunn Lee⁷, Meng-Shiou Lee⁸

Affiliations

¹ Graduate Institute of Biotechnology, National Chung Hsing University, Taichung 40402, Taiwan. just_dfm@hotmail.com.
² Department & Institute of Pharmacology, National Yang-Ming University, Taipei 11221, Taiwan. rich720925@yahoo.com.tw.
³ Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, China Medical University, Taichung 40402, Taiwan. linmk@mail.cmu.edu.tw.
⁴ Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, China Medical University, Taichung 40402, Taiwan. wtchang@mail.cmu.edu.tw.
⁵ Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, China Medical University, Taichung 40402, Taiwan. whpeng@mail.cmu.edu.tw.
⁶ Department of Bioresources, Da-Yeh University, Changhua 51591, Taiwan. fcsun@mail.dyu.edu.tw.
⁷ Management Center, Department of Medical Research and Development, Show Chwan Health Care Sysytem, Changhua 51951, Taiwan. mengshiunn@gmail.com.
⁸ Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, China Medical University, Taichung 40402, Taiwan. leemengshiou@mail.cmu.edu.tw.

Abstract

Taraxacum formosanum (TF) is a medicinal plant used as an important component of health drinks in Taiwan. In this study, a rapid, sensitive and specific loop-mediated isothermal amplification (LAMP) assay for authenticating TF was established. A set of four specific LAMP primers was designed based on the nucleotide sequence of the internal transcribed spacer 2 (ITS2) nuclear ribosomal DNA (nrDNA) of TF. LAMP amplicons were successfully amplified and detected when purified genomic DNA of TF was added in the LAMP reaction under isothermal condition (65 °C) within 45 min. These specific LAMP primers have high specificity and can accurately discriminate Taraxacum formosanum from other adulterant plants; 1 pg of genomic DNA was determined to be the detection limit of the LAMP assay. In conclusion, using this novel approach, TF and its misused plant samples obtained from herbal tea markets were easily identified and discriminated by LAMP assay for quality control.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Base Sequence
DNA Primers / metabolism
DNA, Ribosomal / genetics
DNA, Ribosomal / metabolism
Discriminant Analysis
Genes, Plant*
Molecular Sequence Data
Nucleic Acid Amplification Techniques
Sequence Alignment
Taraxacum / metabolism*
Temperature

Substances

DNA Primers
DNA, Ribosomal