A Species-specific Triplex PCR Assay for Authentication of

Overview

Journal Mitochondrial DNA B Resour

Date 2021 Apr 2

PMID 33796733

Authors

Yuli Zhang

Juan Li

Shiqing Yang

Chao Xu

Roselyn Tehzee Gblinwon

Jianhui Hu

Xiaoxiang Sun

Guohua Xia

Huan Yang

Yuping Shen

Affiliations

Soon will be listed here.

Abstract

A triplex PCR assay was developed to identify animal species and adulteration of a natural medicine (GGEC). Three species-specific primer sets were designed according to the difference in mitochondrial genome of , and . The PCR conditions were optimized and the assay was well validated for high specificity and sensitivity (1 mg/μL). Especially, when artificial adulterants made from the mixture of three species were analyzed, the assay has still exhibited strong capability of differentiation. By using this developed method, two batches out of fourteen commercial GGEC products were identified to be adulterated by . The newly proposed assay showed sufficient merits as a regular tool for the identification of counterfeits or adulterants of GGEC product for their pulverized and processed form, and even Chinese patent medicines composed of these species.

References

Yang H, Yu P, Lu Y, Jiao Z, Chen L, Zhou Y . A novel non-sequencing approach for rapid authentication of Testudinis Carapax et Plastrum and Trionycis Carapax by species-specific primers. R Soc Open Sci. 2018; 5(4):172140. PMC: 5936932. DOI: 10.1098/rsos.172140. View

Girish P, Anjaneyulu A, Viswas K, Shivakumar B, Anand M, Patel M . Meat species identification by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene. Meat Sci. 2011; 70(1):107-12. DOI: 10.1016/j.meatsci.2004.12.004. View

Rodriguez M, Garcia T, Gonzalez I, Asensio L, Mayoral B, Lopez-Calleja I . Identification of goose, mule duck, chicken, turkey, and swine in foie gras by species-specific polymerase chain reaction. J Agric Food Chem. 2003; 51(6):1524-9. DOI: 10.1021/jf025784+. View

Yang H, Zhou Y, Yu P, Yang Y, Jiao Z, Tam J . A novel PCR-based technology for rapid and non-sequencing authentication of Bombyx batryticatus using species-specific primers. Nat Prod Res. 2018; 33(9):1251-1256. DOI: 10.1080/14786419.2018.1466127. View

Yang Y, Zheng Y, Lu B, Jiao Z, Chen L, Gblinwon R . Rapid identification of antlers by species-specific PCR assay. Nat Prod Res. 2019; 34(9):1315-1319. DOI: 10.1080/14786419.2018.1560285. View

Zia Q, Alawami M, Khairil Mokhtar N, Raja Nhari R, Hanish I . Current analytical methods for porcine identification in meat and meat products. Food Chem. 2020; 324:126664. DOI: 10.1016/j.foodchem.2020.126664. View

Wang X, Zhong H, Guo J, Hou F . Morphology and molecular identification of the zoological origin of medicinal seahorses in Chinese herbal markets. Mitochondrial DNA A DNA Mapp Seq Anal. 2020; 31(8):335-345. DOI: 10.1080/24701394.2020.1815719. View

Kitpipit T, Sittichan K, Thanakiatkrai P . Direct-multiplex PCR assay for meat species identification in food products. Food Chem. 2014; 163:77-82. DOI: 10.1016/j.foodchem.2014.04.062. View

Izadpanah M, Mohebali N, Elyasi Gorji Z, Farzaneh P, Vakhshiteh F, Shahzadeh Fazeli S . Simple and fast multiplex PCR method for detection of species origin in meat products. J Food Sci Technol. 2018; 55(2):698-703. PMC: 5785395. DOI: 10.1007/s13197-017-2980-2. View

10.

Azhahianambi P, G J, Gr B, M A, R R, Latha B . Evaluation of multiplex PCR assay for detection of Babesia spp, Ehrlichia canis and Trypanosoma evansi in dogs. Acta Trop. 2018; 188:58-67. DOI: 10.1016/j.actatropica.2018.08.028. View

11.

Chen Y, Yang Y, Qian Y, Gblinwon R, Jiao Z, Chen L . Harnessing multiplex PCR assay targeting specific mitochondrial DNA elements for simultaneous identification of antelope species in . Mitochondrial DNA B Resour. 2020; 4(2):3222-3226. PMC: 7707849. DOI: 10.1080/23802359.2019.1666667. View

12.

Prusakova O, Glukhova X, Afanaseva G, Trizna Y, Nazarova L, Beletsky I . A simple and sensitive two-tube multiplex PCR assay for simultaneous detection of ten meat species. Meat Sci. 2017; 137:34-40. DOI: 10.1016/j.meatsci.2017.10.017. View

13.

Lin L, Zhang Y, Li Y, Fu H, Hu J, Zhou Y . Identification of signature proteins of processed Bombyx batryticatus by comparative proteomic analysis. Int J Biol Macromol. 2020; 153:289-296. DOI: 10.1016/j.ijbiomac.2020.03.010. View

14.

Jiang C, Jin Y, Zhao X, Yuan Y, Zhao Y, Huang L . Rapid and robust authentication of deer antler velvet product by fast PCR-RFLP analysis. Mitochondrial DNA A DNA Mapp Seq Anal. 2017; 29(2):266-272. DOI: 10.1080/24701394.2016.1275599. View

15.

Tasrip N, Desa M, Khairil Mokhtar N, Sajali N, Hashim A, Ali M . Rapid porcine detection in gelatin-based highly processed products using loop mediated isothermal amplification. J Food Sci Technol. 2021; 58(12):4504-4513. PMC: 8478990. DOI: 10.1007/s13197-020-04932-2. View

16.

Yang H, Zheng J, Wang H, Li N, Yang Y, Shen Y . Comparative Proteomic Analysis of Three Gelatinous Chinese Medicines and Their Authentications by Tryptic-digested Peptides Profiling using Matrix-assisted Laser Desorption/Ionization-time of Flight/Time of Flight Mass Spectrometry. Pharmacogn Mag. 2017; 13(52):663-667. PMC: 5701408. DOI: 10.4103/pm.pm_54_17. View