AUTHOR=Zhou Yuxin , Nie Jing , Yu Shiqi , Hu Zhigang , Wang Bo 

TITLE=Multiplex Ligation-Dependent Probe Amplification for Simultaneous Identification of Bungarus multicinctus and Its Common Adulterants in a Single Assay

JOURNAL=Frontiers in Pharmacology

VOLUME=11

YEAR=2020

URL=https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2020.00501

DOI=10.3389/fphar.2020.00501

ISSN=1663-9812

ABSTRACT=<p><italic>Bungarus multicinctus</italic>, an important traditional Chinese medicine, possesses remarkable medicinal activities, while lots of adulterants from other species were misused as <italic>B. multicinctus</italic> for its large demand and resource starvation. In order to accurately identify <italic>B. multicinctus</italic> and its common adulterants such as <italic>Sinonatrix annularis</italic>, <italic>Xenochrophis flavipunctatus</italic>, <italic>Deinagkistrodon acutus</italic>, and <italic>Naja atra</italic>, a simultaneous identification method was designed with multiplex ligation-dependent probe amplification (MLPA) analysis. Five species-specific MLPA probe-couples for <italic>B. multicinctus</italic> and its common adulterants were designed based on the universal primer amplified <italic>COI</italic> sequences, which can specifically detect the five species with no mutual interference, and sensitivity analysis showed as less as 5% <italic>B. multicinctus</italic> or 8.75% adulterants in the mixed samples can be identified in a MLPA assay, especially, the relative quantity of the adulterants can be also inferred based on the MLPA peak area values. Moreover, the results of the present study confirmed the effectiveness of this technique in terms of simultaneous identification of <italic>B. multicinctus</italic> and its common adulterants in an assay, which has great potential for ensuring the safety of this commercially valuable snake species.</p>