Development and validation of DNA sequence-based methods of medicinal herb specie

描述（由申请人提供）：在本提案中，我们概述了一种创新方法，以实现至少两个NCCAMs优先事项：验证用于植物鉴定的DNA方法和开发复杂的DNA认证的植物身份标准参考物质（BI-SRMs）;这将通过帮助确保植物药的质量，安全性和有效性对草药行业产生重大影响。使用DNA进行植物鉴定有许多优点：它可以用于从各种材料类型中识别任何生物，包括原料，粉末和精选加工产品;它非常敏感和强大，甚至可以从几个分子中识别材料;它有能力区分密切相关的物种，杂交种，甚至物种内的谱系;并且具有识别包括有毒或化学惰性掺杂物或添加剂的多于一种主要或污染物质的混合物的能力。在第一阶段，我们有两个具体的目标或目的。在目标1中，我们将通过鉴定两个最佳候选DNA区域来进行预验证研究，以用于以下五个物种中的每一个的鉴定：甘草（甘草）， 毛茛（Hydrastis canadensis）、人参（Panax quinquefolius）、大黄（Rheum palmatum）和黄芩（学名：Escheraria lateriflora）;选择这五个物种是因为它们广泛用于草药，存在已知的掺假问题，并且在分类学上容易混淆，并且可以受益于具有替代形式的植物鉴定。在目标2中，我们将根据AOAC国际定性方法验证指南，对5个物种中的每一个物种的两个基因区域进行完整的单一实验室验证研究，以检测纯样品和不纯样品的特异性和性能。这涉及测试代表五个物种及其密切相关的掺假物的总共200个样本，以及250个混合样本，并计算识别概率（POI）。通过本提案中概述的研究所产生的产品和服务，我们的目标是将DNA认证带到草药行业的最前沿。 公共卫生相关性：在本提案中，我们将完成对五种常用草药（人参、黄精、甘草、大黄和黄芩）的DNA鉴定方法的首次全面验证研究。通过这里生成的数据，我们的目标是将DNA方法带到草药认证的最前沿，使我们不仅能够向公众提供第一个经过验证的DNA测试服务，而且还可以开发第一个经过DNA认证的复杂植物身份标准参考材料，用于替代方法的内部测试和验证研究。这些产品和服务无疑将有助于确保草药产品的安全，质量和功效。

DESCRIPTION (provided by applicant): In this proposal, we outline an innovative approach to accomplish at least two of NCCAMs priorities: validating DNA methods for botanical identification and developing complex DNA-authenticated botanical identity standard reference materials (BI-SRMs); this will have significant impact on the medicinal herb industry by helping to ensure the quality, safety, and efficacy of botanical medicines. There are many advantages to using DNA for botanical authentication: it can be used to identify any organism from a variety of material types, including raw, powder, and select processed products; it is extremely sensitive and powerful and can identify material from even a few molecules; it has ability to discriminate between closely related species, hybrids, and even lineages within species; and has the ability to identify mixtures of more than one primary or contaminating species, including toxic or chemically inert adulterants or additives. In Phase 1 we have two specific aims, or objectives. In Objective 1, we will conduct a pre- validation study by identifying the two best candidate DNA regions for authentication of each of the following five species: licorice (Glycyrrhiza uralensis), goldenseal (Hydrastis canadensis), ginseng (Panax quinquefolius), rhubarb (Rheum palmatum), and skullcap (Scutellaria lateriflora); these five species were chosen because are widely used in herbal medicine, have known issues of adulteration, and taxonomically confusing, and could benefit from having an alternative form of botanical identification. In Objective 2, we will conduc full single laboratory validation studies testing the specificity and performance for two gene regions for each of the five species on pure and impure samples following AOAC Internationals guidelines for qualitative method validation. This involves testing a total of 200 specimens representing the five species and their closely related adulterants, as well as 250 mixed samples, and calculating the Probability of Identification (POI). Through the products and services resulting from the research outlined in this proposal, we aim to bring DNA-authentication to the forefront of the medicinal herb industry. PUBLIC HEALTH RELEVANCE: In this proposal we will complete the first full-validation studies on DNA methods of identification for five commonly used medicinal herbs: ginseng, goldenseal, licorice, rhubarb, and skullcap. Through the data generated here, we aim to bring DNA methods to the forefront of medicinal herb authentication, by allowing us to offer not only the first validated DNA testing services to the public, but also develop the first DNA- authenticated complex botanical identity standard reference materials to use for in-house testing and validation studies of alternative methods. These products and services will undoubtedly help in ensuring the safety, quality, and efficacy of medicinal herb products.