PCR method for identifying bungarus parvus and specific primer thereof

A kind of technology of white flower snake and specific primer, which is applied in the directions of biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., to achieve the effect of shortening the extraction time

CN101613759AActive Publication Date: 2009-12-30华润三九现代中药制药有限公司 +1
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2009-12-30

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Abstract

The invention belongs to the technical field of Chinese medicaments and the identification on Chinese medicinal materials, and particularly relates to a PCR method capable of specifically identifying bungarus parvus and a specific primer thereof. When the method is used to identify the true or false of the bungarus parvus, the identification on the true or false of a sample can be finished through simple DNA extraction, PCR specific amplification and electrophoresis detection. The method has the advantages of simple operation, strong specificity, good repetitiveness and the like. The method is mainly used for the rapid identification of bungarus parvus medicinal materials, preparations containing the bungarus parvus and bungarus parvus living materials.
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Description

technical field

[0001] The invention relates to the technical field of identification of traditional Chinese medicines and Chinese medicinal materials, in particular to a PCR method and specific primers capable of specifically identifying white snakes. It is mainly used for the rapid identification of medicinal materials, preparations containing snakeskin and living materials of snakeskin. Background technique

[0002] The golden snake is a commonly used traditional Chinese medicine recorded in the 2005 edition of the "Chinese Pharmacopoeia", which is derived from the dried body of the Cobra family animal Bungarus multicinctus Blyth (Chinese Pharmacopoeia. One. 2005.151). It has the effects of expelling wind, dredging collaterals and relieving spasm. For rheumatic obstinate numbness, numbness and spasm, hemiplegia, convulsions and convulsions, scrofula malignant sore.

[0003] As an animal medicinal material, Snake chrysalis has definite clinical efficacy, but the commodit...

Examples

Embodiment Construction

[0017] 1 material

[0018] Snake samples came from Hebei Anguo area, Beijing Tongrentang Pharmacy, Beijing Jinxiang Pharmacy, Beijing Sunshine Tongren Pharmacy, Beijing Jinglongtang Pharmacy, etc. See Table 1 for details. The samples were all identified by Director Zhang Ji of the Herbarium of Chinese Medicine Department of China Institute for the Control of Pharmaceutical and Biological Products.

[0019] Table 1 Sample source list

[0020]

[0021]

[0022] 2 Design of specific primers

[0023] Search for the cytochrome b (cytb) sequences of more than 20 species of snakes such as the golden snake and its common confounds such as the golden krait and the red snake in GENBANK, and use DNAMAN to perform homologous comparison, and design a pair of specific primers F1 according to the variable region , F2, the sequence is as follows:

[0024] F1: 5'-GAAATTTCGGCTCTATGCTTATAACCTGTCTTT-3'

[0025] F2: 5'-GGAATCTTATCGATATCTGATTAGTA-3'

[0026] 3DNA extraction

[0027] The...