ABSTRACT
OBJECTIVE: To develop Zaocys dhumnades DNA extraction and detection kit, evaluate the specificity, stability and repeatability of the kit, and investigate the quality of commercial black tip snake products using the kit. METHODS: Using the modern DNA fingerprint technology, the pharmacopoeia Zaocys dhumnades DNA detection method was optimized and improved, and Zaocys dhumnades DNA extraction and detection reagent was developed to obtain the tip snake PCR detection kit. The performance parameters of the kit were evaluated. The kit was used on 18 commercial black tip snake products randomly sampled from Beijing, Tianjin, Changchun and Jilin City.The results were compared with those got by using pharmacopoeia method of DNA test. RESULTS: Zaocys dhumnades DNA testing kit was still valid after repeated freezing and thawing for 5, 10 and 20 times. Among the 18 commercial black tip snake samples 14 were quality goods, and four were adulterants. Repeated experiments of specificity, stability and repeatability displayed exactly the same results. The test result of the kit method was consistent with the pharmacopoeia method. And the kit method could fulfil the DNA testing task in ordinary laboratories. CONCLUSION: Zaocys dhumnades DNA testing kit can identify tip snake products conveniently, accurately and effectively, which can be applied in the rapid detection of Zaocys dhumnades DNA.
ABSTRACT
OBJECTIVE: To develop a kind of Agkistrodon acutus (Günther) DNA test kit, evaluate its quality indexes including specificity, stability, sensitivity and repeatability, and inspect the qualities of commercialAgkistrodon acutus (Günther) samples. METHODS: The Agkistrodon acutus (Günther) DNA test kit was developed and modified according to the method recorded in Chinese Pharmacopoeia (2010 edition). EighteenAgkistrodon acutus (Günther) samples were randomly collected from Beijing, Tianjin, and Changchun. The kit assay was performed to identify these samples with the pharmacopoeia method as the standard control. RESULTS: The kit proved effective after 20 times of freezing and thawing, and repeatability test showed same data for three tests. The lower limit of quantitation was 0.025 g. The specificity test confirmed that 14 samples were genuine, and 4 were adulterants. All of the identification results by the kit assay were in accordance with the ones by the pharmacopoeia method. CONCLUSION: The developed DNA test kit is accurate and effective for identification of Agkistrodon acutus (Günther). Compared with the pharmacopoeia method, it is simpler and more rapid, demonstrating broad prospect in quality inspection of Agkistrodon acutus (Günther).