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Identification of Scolopendra Dispensing Granules by Allele-specific PCR / 中国实验方剂学杂志
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 48-54, 2024.
Article in Chinese | WPRIM | ID: wpr-1005253
ABSTRACT
ObjectiveTo establish an allele-specific polymerase chain reactionPCRmethod for identifying Scolopendra dispensing granules, so as to ensure the quality and therapeutic effects of Scolopendra and its preparations. MethodThe primer interval suitable for the PCR was selected based on the cytochrome c oxidase subunit 3(COX-3) gene sequence of Scolopendra, and the single nucleotide polymorphism (SNP) loci of Scolopendra and its adulterants were mined from the interval for the design of specific primers. The samples of Scolopendra and its adulterants were collected. The PCR system was established and optimized regarding the annealing temperature, cycles, Taq enzymesDNA template amount, PCR instruments, and primer concentrations, and the specificity and applicability of this method were evaluated. ResultThe PCR system was composed of 12.5 μL 2×M5 PCR Mix, 0.4 μL forward primer (10 μmol·L-1), 0.4 μL reverse primer (10 μmol·L-1), 2.5 μL DNA template, and 9.2 μL sterile double distilled water. PCR parameters: Pre-denaturation at 94 ℃ for 3 min, 30 cycles (94 ℃ for 20 s, 62 ℃ for 20 s, 72 ℃ for 45 s), and extension at 72 ℃ for 5 min. After PCR amplification with the system and parameters above, the electrophoresis revealed a bright band at about 135 bp for Scolopendra and no band for the adulterants. ConclusionThe established allele-specific PCR method can accurately identify the medicinal materials, decoction pieces, and standard decoction freeze-dried powder of Scolopendra, as well as the intermediates and final products of Scolopendra dispensing granules, which is of great significance for ensuring the quality and clinical efficacy of Scolopendra and its preparations.

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2024 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2024 Type: Article