目的 对当归不同炮制品中大肠埃希菌变化进行定量分析。方法 建立荧光定量PCR方法，对当归不同炮制品、不同产地、不同收集企业、不同储藏时间的大肠埃希菌进行定量分析。结果 不同炮制品中大肠埃希菌数量变化为生当归 > 土炒当归 > 酒当归；在不同产地的当归中以甘肃岷县地区所含的大肠埃希菌的数量最少；与零售企业相比，生产销售企业的当归和酒当归中大肠埃希菌的数量较少；不同储藏时间对当归和酒当归中大肠埃希菌数量有一定影响，随储藏时间的增加，大肠埃希菌的数量也在增加。对部分代表性样品进行平板计数法与荧光定量PCR法比较时发现，平板计数法结果多呈阴性，荧光定量PCR结果均呈阳性。结论 所建立的荧光定量PCR法特异性、灵敏度、可靠性以及报告周期均优于平板计数法，可为当归不同炮制品中大肠埃希菌的快速、准确定量检测提供有效手段。

Objective To make quantitative analysis for the quantity of Escherichia coli in Angelicae Sinensis Radix (ASR) and its processed products. Methods The fluorescence quantitative PCR method was established to quantitatively analyze E. coli in ASR from different processed products, different producing areas, different enterprises and different storage time. Results The number of E. coli in different processed products was ranked as follows: ASR > ASR stir-frying with soil > ASR stir-frying with wine. And the number of E. coli in the three producing areas of ASR in Min County of Gansu Province was less than that in other producing areas. Compared with the retail enterprises, the number of E. coli in ASR and ASR stir-frying with wine was less in production and sale enterprises. Different storage time had certain effect on the number of E. coli in ASR and ASR stir-frying with wine. With the increase of storage time, the number of E. coli also increased. Plate counting method and fluorescence quantitative PCR method were carried out at the same time for some representative samples. The results showed that the results of the plate counting method were mostly negative, and the results of the fluorescence quantitative PCR were positive. Conclusion The quantitative fluorescence PCR method established in this paper is superior to the plate counting method in specificity, sensitivity, reliability, and reporting cycle, which can provide an effective method for rapid and accurate quantitative detection of E.coli in different processed products of ASR.

R283.6

国家重点研发计划（2017YFC1701902）；国家重点研发计划（2018YFC1707000）；第六批全国老中医药专家学术经验继承（国中医药人教发[2017]29号）