Rapid and reliable distinguish of Morinda officinalis How by loop-mediated isothermal amplification (LAMP) assay

Abstract

The medicinal plant Morinda officinalis How (MO), especially the root, has been frequently used in traditional medicines around the world as an herbal drug for treating variable human disorders and diseases. Various adulterations of MO were found for economic or production limitations. However, authentication of MO from its adulterants by LAMP has not yet been established. The present study introduces a commercially available nucleic acid amplification method, loop-mediated isothermal amplification (LAMP) assay for the distinguish of MO from its adulterants. In this method, we combined DNA barcodes technology to design 2 pairs independent LAMP primers, which based on the internal transcribed spacer 2 (ITS2) sequence of MO’s nuclear ribosomal DNA. Our results showed that the LAMP could amplify the samples as expected and successfully identify target MO, and the limit for DNA template preciseness was verified as 1 × 10− 1 pg/µl. All the visual or real-time turbidity detection was performed within 60 min at approximately 63 °C. The result showed that the LAMP assay and primers we designed have high accuracy and efficiency for the differentiation of MO and its adulterants. Our results illustrated that the proposed low-cost, fast and reliable LAMP assay without the need for expensive equipment or specialized techniques could be a good way for MO rapid authentication.