title: 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.
authors.affiliation: a Department of Clinical Laboratory , Associated Hospital, Beihua University , Jilin , China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Zhang
authors.affiliation: b Department of Molecular Diagnosis , School of Laboratory Medicine, Beihua University , Jilin , China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Zhou
authors.affiliation: c School of Pharmacy, Beihua University , Jilin , China.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Yu
authors.affiliation: b Department of Molecular Diagnosis , School of Laboratory Medicine, Beihua University , Jilin , China.
authors.forename: Jinxia
authors.initials: J
authors.lastname: Ai
authors.affiliation: b Department of Molecular Diagnosis , School of Laboratory Medicine, Beihua University , Jilin , China.
authors.forename: Xuesong
authors.initials: X
authors.lastname: Wang
authors.affiliation: b Department of Molecular Diagnosis , School of Laboratory Medicine, Beihua University , Jilin , China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Gao
authors.affiliation: c School of Pharmacy, Beihua University , Jilin , China.
authors.forename: Guangxin
authors.initials: G
authors.lastname: Yuan
authors.affiliation: b Department of Molecular Diagnosis , School of Laboratory Medicine, Beihua University , Jilin , China.
authors.forename: Mingcheng
authors.initials: M
authors.lastname: Li
mesh_terms: D000818:Animals; D017812:Colubridae / Q000145:classification* / Q000235:genetics; D045303:Cytochromes b / Q000235:genetics; D058893:DNA Barcoding, Taxonomic; D050259:Genes, Mitochondrial; D008516:Medicine, Chinese Traditional
chemical_list: D045303:Cytochromes b
keywords: DNA Kit; PCR; Zaocys dhumnades; bioinformatics