Pcr-based For Gastrodia Elata, Hg²⁺ Detection And A New One-step Visual Detection Of PCR Product By DiSC₂（5）

This paper apply the polymerase chain reaction (PCR) technology for three parts of exploratory research related to the detection metholds with the related substances of traditional Chinese medicine (TCM) as the research object. Chinese Patent medicines is an important outcome of the modernization of Traditional Chinese Medicine, the authenticity and security of Chinese medicinal raw materials are very important for the quality control of CPM. In order to reap huge profits, some merchants often use the similar, similar chemical composition, and the higher content of heavy metals materials for adulteration. The adulteration is very common, especially in some valuable Chinese medicinal raw materials. As the chemotype-driven fingerprints only address the detection of certain compounds, there are some drawbacks if only use the chemical method in the quality control of TCM and CPM. In order to compensate the drawbacks of these chemical detection methods, we apply the PCR technology to the identity the DNA of the adulteration (potato) of Gastrodia elata in Quantianma capsule. What’s more, a heavy metal mercury detection method based on PCR technology was established, which can be used in the detection of heavy metal in Chinese herbal medicines. In order to show the result easily, a new one-step visual detection method was also established. The main work includes three parts as follows:1. We establish a molecular authentication for Gastrodia elata and its possible adulteration in the Chinese Patent medicines. And the adulteration (potato) of Gastrodia elata was detected in Quantianma capsule for the first time. The results prove that the method is specific, simple, reliable and stable, which can successfully detect the adulteration of Gastrodia elata.2. The mercury is an exogenous harmful substances which can affect the security of Chinese medicinal raw materials, and is also harmfui to human health. In order to achieve a quick and easy detection methold, we establish the qualitative and quantitative detection methods by combining the PCR with the two thymine nucleotides on the specific choice of mercury ions.3. PCR product is usually detected by conventional agarose gel electrophoresis, and now has the visual detection method based on G-tetramer. But the previous methods require two steps, which can easily result in contamination of the PCR reaction. So the cyanine dye is used to establish a new one-step visual detection method, it can show the result after the reaction filished immediately. The method has been successfully applied to the detection of target DNA.