title: Mini-SNaPshot multiplex assays authenticate elephant ivory and simultaneously identify the species origin.
abstract: Illegal trading of ivory is mainly responsible for the dramatic decline in elephant populations. Thailand is one of the largest laundering hotspots for African ivory, as the domestic Asian elephant ivory can be legally traded. So, to help combat ivory poaching and smuggling, an efficient method is needed to identify the elephant species from its ivory and ivory products. In this study, a mini-SNaPshot®multiplex assay was developed and fully validated for the identification of confiscated ivory and low DNA template ivory products. Elephantid- and elephant species-specific mitochondrial single nucleotide polymorphisms (SNPs) were identified from 207 mammalian and 1705 elephant/mammoth cytochrome b sequence alignments. Seven informative SNPs were used for assay development. The assay unambiguously and accurately identified authentic elephant ivory and its species of origin on the basis of peak size and color observed in the haplotype profile. The assay was highly efficient for analysis of confiscated ivory and low-template ivory products with a 99.29% success rate (N=140). It was highly reproducible, exhibited no cross-reaction with eight other mammalian DNA; and had 100% identification accuracy. In addition, nested and direct PCR amplification were also compatible with the developed assay. This efficient assay should benefit wildlife forensic laboratories and aid in the prosecution of elephant-related crimes.
authors.affiliation: Forensic Science Program, Department of Applied Science, Faculty of Science, Prince of Songkla University, Thailand. Electronic address: Thitika.k@psu.ac.th.
authors.forename: Thitika
authors.initials: T
authors.lastname: Kitpipit
authors.affiliation: Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Thailand.
authors.forename: Kantima
authors.initials: K
authors.lastname: Thongjued
authors.affiliation: DNP Wildlife Forensic Science Unit, Department of National Parks, Wildlife and Plant Conservation, Thailand.
authors.forename: Kitichaya
authors.initials: K
authors.lastname: Penchart
authors.affiliation: DNP Wildlife Forensic Science Unit, Department of National Parks, Wildlife and Plant Conservation, Thailand.
authors.forename: Kanita
authors.initials: K
authors.lastname: Ouithavon
authors.affiliation: Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University, Thailand.
authors.forename: Wilaiwan
authors.initials: W
authors.lastname: Chotigeat
mesh_terms: D000818:Animals; D003132:Commerce / Q000331:legislation & jurisprudence; D003247:Conservation of Natural Resources / Q000331:legislation & jurisprudence; D003415:Crime / Q000331:legislation & jurisprudence; D045303:Cytochromes b / Q000235:genetics*; D016172:DNA Fingerprinting; D004272:DNA, Mitochondrial / Q000235:genetics*; D004606:Elephants / Q000235:genetics*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D013045:Species Specificity
chemical_list: D004272:DNA, Mitochondrial; D045303:Cytochromes b
keywords: Elephant species; Ivory; Ivory product; SNP typing; SNaPshot multiplex