Identification of Armeniacae Semen Amarum Adulterated in Persicae Semen by Allele-specific Polymerase Chain Reaction
10.13422/j.cnki.syfjx.20210448
- VernacularTitle:位点特异性PCR鉴别桃仁中掺入苦杏仁的方法分析
- Author:
Xue-rui LU
1
;
Zhong-fei SHI
2
;
Bao-xia TENG
2
;
Lin NI
2
;
Ping-rong YANG
2
;
Ping-shun SONG
2
Author Information
1. School of Pharmacy，Lanzhou University，Lanzhou 730000，China
2. Key Laboratory of Quality Control of Chinese Medicinal Materials and Decoction Pieces， National Medical Products Administration，Gansu Institute for Drug Control，Lanzhou 730000，China
- Publication Type:Research Article
- Keywords:
Persicae Semen;
Armeniacae Semen Amarum;
ribosomal DNA internal transcribed spacer （ITS）;
gene sequence;
polymerase chain reaction （PCR）;
single nucleotide polymorphism （SNP） site;
specific primer
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(11):155-161
- CountryChina
- Language:Chinese
-
Abstract:
Objective:Due to the limitation of traditional identification methods of Chinese medicinal materials， the study established a rapid method to identify Persicae Semen mixed with Armeniacae Semen Amarum by allele-specific polymerase chain reaction （PCR）. Method:By comparing the ribosomal DNA internal transcribed spacer （ITS） gene sequences of Persicae Semen and Armeniacae Semen Amarum， single nucleotide polymorphism （SNP） sites were searched and specific primers were designed. Different Persicae Semen and Armeniacae Semen Amarum samples were amplified by PCR， the effects of annealing temperature， primer concentration and cycle number on the PCR reaction system were optimized， and the specificity and detection limit of this method were investigated. In addition， the established PCR method was used to detect the samples of Persicae Semen mixed with different proportion of Armeniacae Semen Amarum from different sources and producing areas. Result:A specific PCR method for identifying Persicae Semen mixed with Armeniacae Semen Amarum was established. When the annealing temperature was 63 ℃ and the number of primer cycles was 30， only Armeniacae Semen Amarum could be amplified with 432 bp specific band， while Persicae Semen samples did not have this band. The minimum detection limit of this method for Armeniacae Semen Amarum was 0.2 ng， and the detection limit for Armeniacae Semen Amarum adulterated in Persicae Semen was 1%. Conclusion:The established allele-specific PCR method can accurately detect whether there is Armeniacae Semen Amarum in Persicae Semen， which can provide experimental basis for the quality control of Persicae Semen and guarantee the safety of its clinical use.
