Abstract
Species identification of unknown biological samples is of fundamental importance for forensic applications, especially in crime detection, poaching, and illegal trade of endangered animals as well as meat fraud. In this study, a novel panel was developed to simultaneously identify 10 different animal species (Gallus domesticus, Anas platyrhynchos domesticus, Ovis aries, Sus scrofa domesticus, Bos taurus, Equus caballus, Columba livia domestica, Rattus norvegicus, Mus musculus, and Canis lupus familiaris) and human beings by amplifying 22 short tandem repeat (STR) loci in a multiplex PCR using a set of five fluorescently labeled dyes. This novel 22-STR panel was validated by optimization of PCR conditions as well as species specificity, sensitivity, reproducibility, precision, DNA mixture, and tissue/organ consistency. The results of developmental validation showed that the 22-STR loci achieved high species specificity among 10 animal species and human beings, and the sensitivity of this panel was 0.09 ng. This 22-STR panel identified different meats in mixed samples, and the minimum detected mixture ratio in the current test was 10% (0.1 ng/1 ng). This sensitive, accurate, and specific 22-STR panel can be used for forensic species identification and the detection of meat fraud and adulteration.
Highlights
Biological samples left behind at crime scenes always contain a great deal of valuable information that can provide helpful clues for the criminal investigations
Species identification of various animal utilizing genetic markers can be applied in the detection of species mislabeling, and the process contributes to food safety by its utilization in meat adulteration cases (Iyengar, 2014)
Before the construction of this novel panel, we amplified each short tandem repeat (STR) locus to evaluate the specificity and amplification efficiency of a pair of primers, and the results showed that specific peaks of each pair of primers were only detected at the corresponding locus for each species, and no peak was found in other species
Summary
Biological samples left behind at crime scenes always contain a great deal of valuable information that can provide helpful clues for the criminal investigations. In addition to human biological specimens, non-human biological samples acquired from a crime scene can suggest certain directions for tracing the suspects. With the continuous progress of biotechnological achievements made in the field of forensic genetics, there has been much interest in the forensic identification of non-human species. Rhinoceros horn and tiger bone were two substances that were formerly used in traditional Chinese medicine. The production and import of these protective animal-derived traditional Chinese medicine are strictly prohibited in China, occasional illegal trade occurs. Species identification is of great importance in the criminal investigation of poaching and illegal trade of endangered animals (Staats et al, 2016). Species identification of various animal utilizing genetic markers can be applied in the detection of species mislabeling, and the process contributes to food safety by its utilization in meat adulteration cases (Iyengar, 2014)
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