Evaluation of DNA Detection Kit for Authenticity of Unibract Fritillary Bulb Based on Pharmacopoeia Method

Abstract

Unibract Fritillary Bulb,as a fomous type of traditional Chinese medicine, derived from Sicuan in China. The objective of the study is to evaluate a kind of DNA assay kit to detect of Unibract Fritillary Bulb from their counterfiets in addition to optimizing its components and protocols. All genomic DNA from 20 samples of Unibract Fritillary Bulb were extracted by Pharmacopoeia method recorded in the expanded edition of China Pharmacopoeia (2010) and assay kit respectively, the PCR technique and Restriction Fragment Length Polymorphism Analysis (RFLP) were carried out to identify the authentication of Unibract Fritillary Bulb. The maximum value of genomic DNA extracted by Pharmacopoeia method is measured as 1.57± 0.05 (OD260/OD280) by Ultraviolet Spectrophotometer, whereas, the value is 1.73±0.10 by assay kit. The results of PCR and RFLP indicated that a single band over 300bp was shown and two distinct bands between 100bp and 250bp in agarose electrophoresis. The data demonstrated that the assay kit was better than the Pharmacopoeia method, especially in extraction quantity and DNA purity of the Unibract Fritillary Bulb nucleic acid; the PCR and RFLP results shown the two methods were consistent compeletiy. The DNA detection Kit for identification of Unibract Fritillary Bulb have good specificity, high sensitivity as well as strong stability, so it is suitable for the rapid detection of Unibract Fritillary Bulb.