Identification of Murrayae Folium et Cacumen by Multiplex Allele-Specific PCR
10.13422/j.cnki.syfjx.20221012
- VernacularTitle:多基原九里香药材的多重位点特异性PCR鉴别
- Author:
Ziyuan CHEN
1
;
Yuyang ZHAO
1
;
Xutao XIE
1
;
Wenbo XIE
2
;
Yan JIN
1
;
Chao JIANG
1
;
Yuan YUAN
1
Author Information
1. 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
2. China Resources Sanjiu Medical & Pharmaceutical Co. Ltd.， Shenzhen 518029， China
- Publication Type:Journal Article
- Keywords:
Murraya exotica;
Murraya paniculata;
multiplex allele-specific polymerase chain reaction （PCR）;
molecular identification
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(17):106-112
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo establish a polymerase chain reaction（PCR） method to accurately discriminate the crude materials of Murrayae Folium et Cacumen， Murraya exotica and M. paniculata. MethodBased on the difference in chloroplast genome sequences of M. exotica and M. paniculata， species-specific identification primers P03 and P04 of M. exotica and M. paniculata were designed according to single nucleotide polymorphism （SNP） on the chloroplast genome. A multiplex allele-specific PCR identification method was established for the identification of M. exotica and M. paniculata following the optimization of annealing temperature， number of cycles， and primer concentration ratio. The established PCR method for identification was explored and verified in terms of tolerance and feasibility by investigating the type of Taq polymerases and PCR system model. ResultIn this multiplex allele-specific PCR identification method， about 330 and 230 bp of specific fragments were amplified from DNA templates of M. exotica and M. paniculata， respectively， under the following conditions：cycle number of 31， annealing temperature of 60 ℃， and primer concentration ratio of P03 and P04 of 1∶2. Consistent results were obtained for samples from different sources. ConclusionThe multiplex allele-specific PCR identification method established in this study can accurately identify the origin of Murrayae Folium et Cacumen， which can be used for the simultaneous identification of M. exotica and M. paniculata by the length of fragments in a single identification assay.
