Identification of Fritillaria taibaiensis and its relatives by real-time PCR with a TaqMan-MGB probe
10.16438/j.0513-4870.2021-0230
- VernacularTitle:TaqMan-MGB实时荧光PCR法检测太白贝母及其近缘种的研究
- Author:
Tian ZHANG
1
;
Jiao CHEN
1
;
Rui-ping JIANG
1
;
Meng ZOU
1
;
Tie-chui YANG
2
;
Shao-bing FU
2
;
Jia-yu ZHOU
1
;
Hai LIAO
1
Author Information
1. School of Life Science and Engineering, Southwest Jiaotong University, Chengdu 610031, China
2. Qinghai Lvkang Biological Development Co., Ltd., Xining 810003, China
- Publication Type:Research Article
- Keywords:
TaqMan-MGB probe;
italic>Fritillaria taipaiensis;
ITS1;
real-time PCR;
molecular identification
- From:
Acta Pharmaceutica Sinica
2021;56(9):2577-2583
- CountryChina
- Language:Chinese
-
Abstract:
The molecular identification of Fritillaria taibaiensis and its relatives was studied by real-time PCR with a TaqMan-MGB probe. DNA was extracted from F. taibaiensis and its relatives. According to the sequence of ITS1 region, the mutation sites of F. taipaiensis and its related species were identified by MEGA7.0 software. The specific primers (a pair) and a TaqMan-MGB probe were designed by Primer Premier 6.0 software. In the Roche LightCycler 96 system, the lowest limit of detection for F. taipaiensis DNA template was 0.002 39 ng·μL-1, and the optimal Tm value range was 60 and 61 ℃. Specificity identification showed that the method had good specificity for F. taipaiensis, as it could be distinguished from other 13 different Fritillaria species including F. unibracteata. Since this method could accurately identify F. taipaiensis and its related species, it provides technical support for rational development of F. taipaiensis resources, management of Chinese medicinal market and supervision of raw materials in Chinese medicine manufacturing enterprises.
