Detection of 400-year-old Yersinia pestis DNA in human dental pulp: An approach to the diagnosis of ancient septicemia

Abstract

Ancient septicemic plague epidemics were reported to have killed millions of people for 2 millenniums. However, confident diagnosis of ancient septicemia solely on the basis of historical clinical observations is not possible. The lack of suitable infected material has prevented direct demonstration of ancient septicemia; thus, the history of most infections such as plague remains hypothetical. The durability of dental pulp, together with its natural sterility, makes it a suitable material on which to base such research. We hypothesized that it would be a lasting refuge for Yersinia pestis, the plague agent. DNA extracts were made from the dental pulp of 12 unerupted teeth extracted from skeletons excavated from 16th and 18th century French graves of persons thought to have died of plague (''plague teeth'') and from 7 ancient negative control teeth. PCRs incorporating ancient DNA extracts and primers specific for the human ␤-globin gene demonstrated the absence of inhibitors in these preparations. The incorporation of primers specific for Y. pestis rpoB (the RNA polymerase ␤-subunit-encoding gene) and the recognized virulence-associated pla (the plasminogen activatorencoding gene) repeatedly yielded products that had a nucleotide sequence indistinguishable from that of modern day isolates of the bacterium. The specific pla sequence was obtained from 6 of 12 plague skeleton teeth but 0 of 7 negative controls (P < 0.034, Fisher exact test). A nucleic acid-based confirmation of ancient plague was achieved for historically identified victims, and we have confirmed the presence of the disease at the end of 16th century in France. Dental pulp is an attractive target in the quest to determine the etiology of septicemic illnesses detected in ancient corpses. Molecular techniques could be applied to this material to resolve historical outbreaks.

Molecular analysis of skeletal tuberculosis in an ancient Egyptian population

“…In general, this approach has been used for the detection of tuberculosis [1±10], and also for the identi®cation of other pathogens, such as M. leprae [22,23], Plasmodium falciparum [24], Trypanosoma cruzi [25], Yersinia pestis [26] and Escherichia coli [27]. Previous studies had shown that this approach is applicable to ancient Egyptian material, as M. tuberculosis was identi®ed in one of the mummies investigated [5].…”

Absence of Yersinia pestis-specific DNA in human teeth from five European excavations of putative plague victims

“…Whether these historically described diseases correspond to bubonic plague, whose aetiological agent was identified to be the bacterium now known as Yersinia pestis by Alexandre Yersin in 1894 (Yersin, 1894), has been the subject of controversy (Cohn, 2002;Scott & Duncan, 2001). Recently, DNA specific for Y. pestis was amplified from 16th and 18th century human teeth believed to be from French plague victims (Drancourt et al, 1998) and 14th century French Black Death victims (Raoult et al, 2000). The lead authors of these reports believe that the consideration of any cause for the Black Death other than Y. pestis is now speculative .…”

Plagued by doubt and viral misinformation: the need for evidence-based use of historical disease images

“…[38][39][40] Since the first publication announcing ancient human dental pulp containing Yersinia pestis ancient DNA, 41 scientists have reconstructed its full genome, 42 reconstructed its evolutionary history, 43 and suggested the potential endemicity of the plague in Europe 44, 45 and the Middle East, 46 all of which open new historical questions. Other studies have also broadened the historical narrative in various ways: adding more whole genomes to the pool of Y pestis data, 47 highlighting the variety of potential Y pestis hosts and vectors 48,49 and the role of microclimates in pathogen survival, 50 exploring immunological responses, 51 assessing mortality patterns, 52 and suggesting a potential route and impact of the plague in historical sub-Saharan Africa.…”