聚合酶链式反应-限制性内切酶多态法检查川贝母的掺伪情况

目的应用DNA分子鉴定技术检测川贝母的掺伪情况。方法收集111份川贝母及其主要伪品对口药材,应用聚合酶链式反应-限制性内切酶多态法(PCR-RPLF)对川贝母、川贝母伪品及混合样品进行检测。结果该方法能够检出川贝母中伪品的掺伪量,随着掺伪量的增加,未切DNA条带灰度增强而被切条带减弱;未切条带相对于被切条带而言,其灰度比率相应增加;当掺伪量达75%时,几乎观察不到被切条带。结论所用方法灵敏度高,通过测量琼脂糖凝胶电泳图中DNA条带的灰度值强弱,可知道川贝母中大概的掺伪比例,可用于川贝母样品中掺伪量的检测。

Research on the adulterants identification in Fritillariae cirrhosae Bulbus with PCR-RPLF method

OBJECTIVE To distinguish the common adulterants in Fritillariae cirrhosae Bulbus by the polymerase chain reaction-restriction fragment length polymorphisms(PCR-RFLP)method.METHODS 111 different kinds of the F.cirrhosae Bulbus from different places were collected.PCR-RPLF technology was used to distinguish the F.cirrhosae Bulbus and the mixed powders,and then the percentage content of adulterants was calculated.RESULTS 1%adulteration in F.cirrhosae Bulbus can be detected by PCR-RPLF technology.With the increase of adulteration amount,the uncutted DNA bands became stronger and the cutted DNA bands became weaker.The grayscale ratio of the uncutted bands to that of the cutted increased accordingly.When the proportion of the adulterants increased to 75%,there were nearly no visible cutted DNA bands.CONCLUSION The PCR-RPLF technology is sensitive for the identification of the adulterants in F.cirrhosae Bulbus.Furthermore,it is possible to know the approximate proportion of the adulterants by measuring the grayscale of the DNA bands on the map of agarose gel electrophoresis.