道地中药材光慈菇检测试剂盒的研制和评价

doi:10.3969/j.issn.1674-7968.2022.06.015

Abstract
Figure/Table
References (16)
Related Citation (15)

Download: PDF (5128 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract In recent years, the application of traditional Chinese medicine has become more and more popular, and the market demand is increasing, but the mixed phenomenon of counterfeits is serious. The development of identification kits based on DNA detection technology is of significant practical significance for this medicinal material. This study was based on the phylogenetic tree constructed in early stage of the experiment on A. edulis and its fakes, and specific PCR primers for A. edulis were designed on the basis of internal transcribed spacers (ITS) sequence analysis to optimize the use of specific detection reagents and reaction conditions. According to the evaluation criteria of in vitro diagnostic kits, a test kit was developed for A. edulis, and the test kit was evaluated for specificity, detection limit, repeatability, shelf life and stability. According to the test results, the kit had good specificity, the detection limit was 0.03 μg/μL, and excellent stability. The specific identification kit of authentic Chinese medicinal material A. edulis was accurate, fast and effective for the identification of A. edulis, and has broad application prospects.

Key words： Amana edulis PCR Detection kit Molecular identification Nucleic acid dye SYBR Green Ⅰ

Received: 01 October 2021

ZTFLH:

S567.239

Corresponding Authors: * zhaoqun626@163.com

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	SUN Chuan-Bo
	HE Yan-Fei
	ZHANG Jing
	WANG Yu-Wei
	ZHAO Qun

Cite this article:

SUN Chuan-Bo,HE Yan-Fei,ZHANG Jing, et al. Development and Evaluation of the Detection Kit for Authentic Chinese Medicinal Materials Bulb of Edible Tulip[J]. 农业生物技术学报, 2022, 30(6): 1202-1209.

URL:

http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2022.06.015 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2022/V30/I6/1202

[1] 陈随清, 王利丽. 2003. rRNA基因序列分析在中药品种鉴定中的应用及研究进展[J]. 中医学报, 18(105): 86-88.
(Chen S Q, Wang L L.2003. Application and development of rRNA sequencing in origin identification of Chinese materia medica[J]. Acta Chinese Medicine, 18(105): 86-88.)
[2] 程龙, 葛斐林, 刘泽邦, 等. 2015. 中药资源DNA条形码鉴定方法研究进展[C]//, 北京: 第四届中国中药商品学术大会暨中药鉴定学科教学改革与教材建设研讨会论文集, pp. 398.
(Cheng L, Ge F L, Liu Z B, et al.2015. Research progress of DNA bar code identification methods of traditional Chinese medicine resources[C]//, Beijing: Proceedings of the 4th China traditional Chinese medicine commodity academic conference and seminar on teaching reform and textbook construction of traditional Chinese medicine identification, pp. 398.)
[3] 侯飞侠, 曹静, 王沙沙, 等. 2017. 冬虫夏草PCR快速检测试剂盒的研制与效果评价[J]. 中国中药杂志, 42(6):1125-1129.
(Hou F X, Cao J, Wang S S, et al.2017. Development and effect evaluation of Cordyceps sinensis PCR rapid detection kit[J]. China Journal of Chinese Materia Medica, 42(6): 1125-1129.)
[4] 黄璐琦, 袁媛. 2014. 中药分子鉴定操作指南[M].上海科学技术出版社, 上海. pp. 49.
(Huang L Q, Yuan Y.2014. Operation Guide for Molecular Identification of Traditional Chinese Medicine[M]. Shanghai: Shanghai Science and Technology Press, China. pp. 49.)
[5] 江苏新医学院. 1977. 中药大辞典(下册)[M].上海人民出版社, 上海. pp. 2713.
(Jiangsu New Medical College. 1977. Dictionary of traditional Chinese medicine (Volume Ⅱ)[M]. Shanghai People's publishing house, Shanghai, China. pp. 2713.)
[6] 刘邦强, 王业发, 余淑芳, 等. 2005. 同名异科慈菇真伪品鉴别比较[J]. 中国民族民间医药杂志, 12(73):111-112.
(Liu B Q, Wang Y F, Yu S F, et al.2005. Identification and comparison of true and false products of different families of Amana edulis[J]. Comparison on Identification of Cigu from Different Families, 12(73): 111-112.)
[7] 刘婷婷, 于栋华, 刘树民. 2020. 山慈菇的本草考证及现代研究进展[J]. 中国药房, 31(24):3055-3059.
(Liu T T, Yu D H, Liu S M.2020. Herbal textual research and modern research progress of Amana edulis[J]. China Pharmacy, 31(24): 3055-3059.)
[8] 童静玲, 王银萍, 朱让腾. 2011. 山慈菇与光慈菇的鉴别及正确使用[J]. 海峡药学, 23(2): 28-29.
(Tong J L, Wang Y P, Zhu R T.2011. Identification and correct use of Amana edulis and bulb of edible tulip[J]. Strait Pharmaceutical Journal , 23(2): 28-29.)
[9] 王晓阁, 龙全江, 赵悦. 2015. 光慈菇药用历史变迁与现代研究概况[J]. 甘肃中医学院学报, 32(6): 72-74.
(Wang X G, Long Q J, Zhao Y.2015. Overview of medicinal historical changes and modern research of Amana edulis[J]. Journal of Gansu University of Chinese Medicine, 32(6): 72-74.)
[10] 吴文如, 安鑫, 来慧丽, 等. 2019. 基于ITS2序列位点特异性PCR鉴别广藿香及其混伪品[J]. 时珍国医国药, 30(4): 897-900.
(Wu W R, An X, Lai H L, et al.2019. Identification of Pogostemon cablin (Blanco) Benth. with the counterfeit products by allele-specific PCR based on ITS2 sequence[J]. Lishizhen Medicine and Materia Medica Research, 30(4): 897-900.)
[11] 吴正军, 朱再标, 郭巧生, 等. 2012. 老鸦瓣种子生理及其萌发特性研究[J]. 中国中药杂志, 37(5):575-579.
(Wu Z J, Zhu Z B, Guo Q S, et al.2012. Study on physiological and germination characteristics of Tulipa edulis seed[J]. China Journal of Chinese Materia Medica, 37(5): 575-579.)
[12] 肖明阳, 罗云清, 王志成. 2021. rDNA-ITS序列分析方法在药用植物研究中的应用[J/OL]. 分子植物育种: 1-16. http://kns.cnki.net/kcms/detail/46.1068.S.20210209.1651.013.html.
[13] 于文静, 周亭亭, 张丽华, 等. 2014. 蕲蛇DNA检测试剂盒的评价与应用[J].中国药学杂志, 49(11): 999-1003.
(Yu W J, Zhou T T, Zhang L H, et al.2014. Evaluation and application of Agkistrodon acutus DNA detection kit[J]. Chinese Pharmaceutical Journal, 49(11): 999-1003.)
[14] 赵群, 刘枫, 宋向文, 等. 2017. 光慈菇药材的特异性PCR鉴定[J]. 中药材, 40(7): 1540-1546.
(Zhao Q, Liu F, Song X W, et al.2017. Specific PCR identification between Amana edulis and its adulterants[J]. Journal of Chinese Medicinal Materials, 40(7): 1540-1546.)
[15] Chen S, Yao H, Han J, et al.2010. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species[J]. PLOS ONE, 5(1): e8613.
[16] Tamura K, Dudley J, Nei M, et al.2007. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0[J]. Molecular Biology and Evolution, 24(8): 1596-1599.