Development and evaluation of a PCR-based assay kit for authentication of Zaocys dhumnades in traditional Chinese medicine

Abstract

We developed a kind of Zaocys dhumnades DNA test kit and it’s indexes including specificity, sensitivity and stability were evaluated and compared with the method recorded in Chinese Pharmacopoeia (2010 edition). The bioinformatics technology was used to design primers, sequencing and blast, in conjunction with PCR technology based on the characteristics of Z. dhumnades cytochrome b (Cyt b) gene. The efficiency of nucleic acid extraction by the kit was done in accordance with Pharmacopoeia method. The kit stability results proved effective after repeated freezing and thawing 20 times. The sensitivity results indicated that the lowest amount detected by the kit was 0. 025 g of each specimen. The specificity test of the kit was 100% specific. All repeatability tests indicated the same results when conducted three times. Compared with the method recorded in Chinese Pharmacopoeia, the PCR-based assay kit by our team developed is accurate, effective in identification of Z. dhumnades, it is simple and fast, demonstrating a broad prospect in quality inspection of Z. dhumnades in the future.

Keywords:

• Zaocys dhumnades
• DNA Kit
• PCR
• bioinformatics

Disclosure statement

The authors declare that they have no competing interests. The authors alone are responsible for the content and writing of the paper.

Additional information

Funding

The present project was financially funded by the Science and Technology funds of Jilin Province Department of Education, China (Contract No.: 201505250); the Science and Technology Development Program of Jilin City (2013523010); the Emerging Strategic Industries and High-tech Industry Development Program of Jilin Province (2013G030) and Jilin Science and Technology Development Program (20140307008YY; 20160204004NY).