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.
Highlights
The loop-mediated isothermal amplification (LAMP) assay is a well-developed method capable of rapid and specific DNA amplification
To design the LAMP primers for the authentication of Taraxacum formosanum (TF), the internal transcribed spacer 2 (ITS2) regions of the nuclear ribosomal DNA of TF was used as a target DNA region
The interspecies variation of ITS2 among TF and its adulterants ranged from 27.02% to 59.45%, which is very useful when designing LAMP primers
Summary
The loop-mediated isothermal amplification (LAMP) assay is a well-developed method capable of rapid and specific DNA amplification. Because of the strand displacement activity of Bst DNA polymerase, the LAMP reaction can be performed without the denaturation step of PCR [1,2]. The LAMP method has been applied to the detection of various microbes or pathogens such as protozoa, bacteria, and viruses [3,4,5,6,7,8,9,10]. LAMP is time-effective, with high specificity and sensitivity and requires relatively simple equipment; it has become a very powerful tool for developing a nucleic acid-based diagnostic method for identification or authentication of bio-resources
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