婴儿健脾散中川贝母荧光PCR检测方法的建立及应用

Establishment and Application of Real-time Fluorescence PCR Detection of Fritillaria Cirrhosae Bulbus in Ying’er Jianpi Powder

关键词:  川贝母  双重实时荧光PCR  婴儿健脾散  真伪

Key words:  Fritillaria Cirrhosae Bulbus  dual real-time fluorescence PCR  Ying'er Jianpi powder  authenticity

摘要:

目的 探讨双重实时荧光PCR检测方法对婴儿健脾散中川贝母成分真伪鉴别的可行性。方法 使用特异性TaqMan引物和探针建立双重荧光PCR检测法,对川贝母及其常见伪品进行鉴别,并对其灵敏度、特异性和自制模拟婴儿健脾散处方进行评估。进一步使用该方法对市场流通的婴儿健脾散中的川贝母真伪进行检测。结果 建立的双重实时荧光PCR检测方法特异性高,灵敏度达0.000 1 ng·µL^-1;对49批婴儿健脾散样品抽查检出6批不合格,其中2批未检出贝母属成分,4批检出贝母属成分但未检出川贝母成分。结论 本双重实时荧光PCR法可成功鉴定婴儿健脾散中川贝母的真伪,可为含川贝母成分复方制剂的川贝母真伪鉴别提供依据。

Abstract:

OBJECTIVE To explore the feasibility of dual real-time fluorescence PCR in the identification of the authenticity of Fritillaria Cirrhosae Bulbus in Ying'er Jianpi powder. METHODS A dual detection method was established by using specific TaqMan primers and probes to identify Fritillaria Cirrhosae Bulbus and its common counterfeits. Then, its sensitivity, specificity and self-made formula of simulated Ying'er Jianpi powder were evaluated. The method was further used to detect the authenticity of Fritillaria Cirrhosae Bulbus in Ying'er Jianpi powder circulating in the market. RESULTS The established dual real-time fluorescent PCR method had high specificity with a sensitivity of 0.000 1 ng·µL^-1. Six of 49 batches of Ying'er Jianpi powder samples were found to be unqualified, including 2 batches detected no Fritillaria L. and 4 batches detected no Fritillaria Cirrhosae Bulbus. CONCLUSION This dual real-time fluorescence PCR method can successfully identify the authenticity of Fritillaria Cirrhosae Bulbus in Ying'er Jianpi powder, and can provide a basis for the identification of Fritillaria Cirrhosae Bulbus in compound preparations.