Environmental DNA as an innovative technique to identify the origins of falsified antimalarial tablets—a pilot study of the pharmabiome

Abstract

Falsified medicines are a major threat to global health. Antimalarial drugs have been particularly targeted by criminals. As DNA analysis has revolutionized forensic criminology, we hypothesized that these techniques could also be used to investigate the origins of falsified medicines. Medicines may contain diverse adventitious biological contamination, and the sealed nature of blister-packages may capture and preserve genetic signals from the manufacturing processes allowing identification of production source(s). We conducted a blinded pilot study to determine if such environmental DNA (eDNA) could be detected in eleven samples of falsified and genuine artesunate antimalarial tablets, collected in SE Asia, which could be indicative of origin. Massively Parallel Sequencing (MPS) was used to characterize microbial and eukaryote diversity. Two mitochondrial DNA analysis approaches were explored to detect the presence of human DNA. Trace eDNA from these low biomass samples demonstrated sample specific signals using two target markers. Significant differences in bacterial and eukaryote DNA community structures were observed between genuine and falsified tablets and between different packaging types of falsified artesunate. Human DNA, which was indicative of likely east Asian ancestry, was found in falsified tablets. This pilot study of the ‘pharmabiome’ shows the potential of environmental DNA as a powerful forensic tool to assist with the identification of the environments, and hence location and timing, of the source and manufacture of falsified medicines, establish links between seizures and complement existing tools to build a more complete picture of criminal trade routes. The finding of human DNA in tablets raises important ethical issues that need to be addressed.

Soil and Sediments in Natural Underground Ecosystems as a Source of Culturable Micromycetes: A Case Study of the Brestovská Cave (Western Tatras, Slovakia)

“…Four isolates of Mucormycota phylum were obtained from the soil and sediment of the Brestovská Cave. Ambomucor seriatoinflatus was previously isolated from soil samples from Mongolia, China, and Alaska (with low prevalence); however, this is its first occurrence in the cave ecosystems [97,98]. Moreover, three species of the Mortierella genus were found in our study.…”

The uncertain role of substandard and falsified medicines in the emergence and spread of antimicrobial resistance

“…A final important aspect is improving our understanding of the manufacture and trade of falsified medicines to inform and prioritize actions and interventions to reduce their occurrence. Part of this could involve innovative forensic approaches using environmental DNA and stable isotope analysis to understand the origins of falsified medicines 67 , 68 . Social network analysis could then help identify common trade routes and manufacturing hotspots, and hence where interventions might be most effectively targeted 69 .…”

Forensic investigation of falsified antimalarials using isotope ratio mass spectrometry: a pilot investigation

“…Pollen and calcium carbonate analyses were important in providing evidence that falsified oral antimalarial artesunate—in a large epidemic that afflicted mainland Southeast Asia in the late 1990s and early 2000s—was from southern China 6 . More recently, environmental DNA has been shown to offer promise for providing signatures, specific for time and place, for the ingredients and manufacturing sites of falsified medicines 7 . There has been much more research and significant recent innovations in the forensic investigation of the illegal wildlife trade to provide evidence, for instance, on the habitat of elephant victims of the ivory trade.…”