Search

Chemical decomposition and fragmentation may limit the detection of ancient host and microbial DNA while some proteins can be detected for extended periods of time. We applied paleoproteomics on 300-year-old dental pulp specimens recovered from 16 individuals in two archeological funeral sites in France, comprising one documented plague site and one documented plague-negative site. The dental pulp paleoproteome of the 16 teeth comprised 439 peptides representative of 30 proteins of human origin and 211 peptides representative of 27 proteins of non-human origin. Human proteins consisted of conjunctive tissue and blood proteins including IgA immunoglobulins. Four peptides were indicative of three presumable Yersinia pestis proteins detected in 3/8 dental pulp specimens from the plague-positive site but not in the eight dental pulp specimens collected in the plague-negative site. Paleoproteomics applied to the dental pulp is a new and innovative approach to screen ancient individuals for the detection of blood-borne pathogens and host inflammatory response.

Background: The cause of past plague pandemics was controversial but several research teams used PCR techniques and dental pulp as the primary material to reveal that they were caused by Yersinia pestis. However, the degradation of DNA limits the ability to detect ancient infections., Methods: We used for the first time immuno-PCR to detect Yersinia pestis antigens; it can detect protein concentrations 70 times lower than the standard ELISA. After determining the cut-off value, we tested 34 teeth that were obtained from mass graves of plague, and compared previous PCR results with ELISA and immuno-PCR results., Results: The immuno-PCR technique was the most sensitive (14 out of 34) followed by the PCR technique (10 out of 34) and ELISA (3 out of 34). The combination of these three methods identified 18 out of 34 (53%) teeth as presumably being from people with the plague., Conclusion: Immuno-PCR is specific (no false-positive samples were found) and more sensitive than the currently used method to detect antigens of ancient infections in dental pulp. The combination of three methods, ELISA, PCR and immuno-PCR, increased the capacity to identify ancient pathogens in dental pulp.

Background: Historical records suggest that multiple burial sites from the 14th-16th centuries in Venice, Italy, were used during the Black Death and subsequent plague epidemics., Methodology/principal Findings: High throughput, multiplexed real-time PCR detected DNA of seven highly transmissible pathogens in 173 dental pulp specimens collected from 46 graves. Bartonella quintana DNA was identified in five (2.9%) samples, including three from the 16th century and two from the 15th century, and Yersinia pestis DNA was detected in three (1.7%) samples, including two from the 14th century and one from the 16th century. Partial glpD gene sequencing indicated that the detected Y. pestis was the Orientalis biotype., Conclusions: These data document for the first time successive plague epidemics in the medieval European city where quarantine was first instituted in the 14th century.

From AD 1347 to AD 1353, the Black Death killed tens of millions of people in Europe, leaving misery and devastation in its wake, with successive epidemics ravaging the continent until the 18(th) century. The etiology of this disease has remained highly controversial, ranging from claims based on genetics and the historical descriptions of symptoms that it was caused by Yersinia pestis to conclusions that it must have been caused by other pathogens. It has also been disputed whether plague had the same etiology in northern and southern Europe. Here we identified DNA and protein signatures specific for Y. pestis in human skeletons from mass graves in northern, central and southern Europe that were associated archaeologically with the Black Death and subsequent resurgences. We confirm that Y. pestis caused the Black Death and later epidemics on the entire European continent over the course of four centuries. Furthermore, on the basis of 17 single nucleotide polymorphisms plus the absence of a deletion in glpD gene, our aDNA results identified two previously unknown but related clades of Y. pestis associated with distinct medieval mass graves. These findings suggest that plague was imported to Europe on two or more occasions, each following a distinct route. These two clades are ancestral to modern isolates of Y. pestis biovars Orientalis and Medievalis. Our results clarify the etiology of the Black Death and provide a paradigm for a detailed historical reconstruction of the infection routes followed by this disease.

A rapid diagnostic dipstick test (RDT) that detects Yersinia pestis F1 antigen has been recently applied on 18 putative plague victims exhumed from four archaeological burial sites in southeastern France dating back to the 16(th), 17(th) and 18(th) centuries. The Y. pestis antigen F1 was detected in 12 ancient samples out of 18 (67%). Negative controls confirmed their negativity (100%). Our results emphasize that the detection threshold of the RDT for plague (0.5 ng/ml) is sufficient for a first retrospective diagnosis of Y. pestis infection in ancient remains, and confirm the high specificity and sensitivity of the assay. Double-blind analyses performed by using two different techniques (RDT and 'suicide PCR') led us to the identification of the Y. pestis F1 antigen and the Y. pestis pla and gplD genes. These data provide clear evidence of the presence of Y. pestis in the examined specimens.

Yersinia pestis DNA was recently detected in human remains from 2 ancient plague pandemics in France and Germany. We have now sequenced Y pestis glpD gene in such remains, showing a 93-bp deletion specific for biotype Orientalis. These data show that only Orientalis type caused the 3 plague pandemics.

The article discusses the history of the bubonic plague, especially in Europe. In particular it discusses methods of examining mass burials associated with the disease, especially in early modern France. Among other topics addressed are the isolation of the associated bacteria by French bacteriologist Alexandre Yersin, history of cases of the disease, and attempts to create a vaccine.

The article examines the role of Yersinia pestis in past pandemic diseases by applying a rapid diagnostic test that detects the bacterium's F1 antigen to the remains of 28 putative plague victims that are exhumed from several 16th- to 18th-century burial sites in southeastern France. Also cited are the issues' implications for physical anthropology.

Abstract: The skeletons of four Benedictine nuns and two priests were exhumed from two burial sites in central France. The corpses were dated to the 16th– 18th centuries (archaeological dating). Thick layers of lime were associated with their burials. During the Second Plague Pandemic, lime was associated with plague burials. We hypothesize that Yersinia pestis infection might have been the cause of death of these religious. A rapid diagnostic test for plague (RDT), that detects Y. pestis F1 antigen, was applied to all six putative plague victims. Y. pestis F1 antigen was detected in all six skeletons (100%). The F1 antigen concentrations ranged from 2.5 to 0.625ng/ml. The eight samples used as negative controls yielded negative results. Soils samples taken from archaeological sites related to both positive and negative samples tested negative for F1 antigen. In earlier reports we provided suggestive evidence that the RDT for plague might be diagnostic for the detection of Yersinia pestis in ancient human remains. This putative evidence was based on four French and one German reputed plague sites in which RDT for plague and PCR sequenced-based analyses were performed. Both tests gave results consistent with our interpretation that RDT is effective in identifying plague in skeletal remains. Several different European putative plague burial sites are currently under investigation. We hope to obtain additional material to allow statistical validation of our results obtained thus far. We now confirm that the detection threshold of the RDT for plague (0.5ng/ml) is sufficient for a retrospective diagnosis of plague in ancient skeletal remains. This method supplements the historical record with independent biologic evidence. Thus, our hypothesis that the six religious were infected by Y. pestis during the plague outbreaks which occurred in the two sites, Poitiers and La Chaize-le-Vicomte, between 1587 and 1632, has independent biologic support. [Copyright &y& Elsevier]

Abstract: A rapid diagnostic dipstick test (RDT) that detects Yersinia pestis F1 antigen has been recently applied on 18 putative plague victims exhumed from four archaeological burial sites in southeastern France dating back to the 16th, 17th and 18th centuries. The Y. pestis antigen F1 was detected in 12 ancient samples out of 18 (67%). Negative controls confirmed their negativity (100%). Our results emphasize that the detection threshold of the RDT for plague (0.5 ng/ml) is sufficient for a first retrospective diagnosis of Y. pestis infection in ancient remains, and confirm the high specificity and sensitivity of the assay. Double-blind analyses performed by using two different techniques (RDT and ‘suicide PCR’) led us to the identification of the Y. pestis F1 antigen and the Y. pestis pla and gplD genes. These data provide clear evidence of the presence of Y. pestis in the examined specimens. To cite this article: R. Bianucci et al., C. R. Biologies 330 (2007). [Copyright &y& Elsevier]