Identification of Pheretima （Pheretima aspergillum） Formula Granule by Allele-specific PCR
10.13422/j.cnki.syfjx.20221211
- VernacularTitle:地龙（参环毛蚓）配方颗粒的位点特异性PCR鉴别
- Author:
Li HU
1
;
Yuyang ZHAO
2
;
Yuan YUAN
2
;
Chao JIANG
2
Author Information
1. School of Pharmacy， Anhui University of Traditional Chinese Medicine， Hefei 230012， China
2. State Key Laboratory Breeding Base of Dao-Di Herbs， National Resource Center for Chinese Materia Medica，China Academy of Chinese Medical Sciences，Beijing 100700，China
- Publication Type:Journal Article
- Keywords:
P. aspergillum;
formula granule;
polymerase chain reaction （PCR）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(17):119-126
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveIn order to ensure the safety and effectiveness of clinical drug use ， the identification method of mixed and adulterated specific polymerase chain reaction （PCR） identification of Pheretima aspergillum and its processed products was established. MethodBased on the cytochrome C oxidase subunit I sequence of P. aspergillum， primers were designed to cover the whole sequences， and the stable DNA ranges suitable for the identification of Pheretima （P. aspergillum） formula granule were screened out. Specific primers were designed according to the specific single nucleotide polymorphisms （SNP） of P. aspergillum in the stable DNA range. The P. aspergillum and its mixture were collected respectively， the PCR reaction system was established and optimized， and PCR reaction system and procedure were optimized， and the tolerance and applicability were investigated. ResultWhen the annealing temperature was 62 ℃ and the cycle number was 36， both P. aspergillum formula granule and its formula particles could amplify a single specific identification band of about 170 bp， and the other 20 adulterants and negative controls had no band. ConclusionThe allele-specific PCR identification method established in this study can quickly and accurately identify the P. aspergillum formula granule. The orgin of Chinese herbal medicine and decoction pieces and P. aspergillum were accurately identified. It can also provide a reference for other studies on the quality standard research of other Chinese herbal formula granule.
