Emerg Infect DisEmerging Infect. DisEIDEmerging Infectious Diseases1080-60401080-6059Centers for Disease Control and Prevention21529431333816210-177710.3201/eid1705.101777Letters to the EditorLetterYersinia pestis DNA Sequences in Late Medieval Skeletal Finds, BavariaYersinia pestis DNA SequencesTranThi-Nguyen-NyRaoultDidierDrancourtMichelAuthor affiliation: Université de la Méditerranée, Marseille, FranceAddress for correspondence: Michel Drancourt, Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes, Unité de Mixte de Recherche, Centre National de la Recherche Scientifi que, 6236 –Institut de Recherche et de Développement 198, Faculté de Médecine, 27 Blvd Jean Moulin, 13385 Marseille CEDEX 5, France; email: michel.drancourt@univmed.fr52011175955957WiechmannI , HarbeckM , GrupeG . Yersinia pestis DNA sequences in late medieval skeletal finds, Bavaria. Emerg Infect Dis. 2010;17:1806–7.Keywords: plagueYersinia pestisBlack Deathgenotypinghomopolymeric tractbacterialetter

To the Editor: We read with interest the report by Wiechmann et al. that, in the investigation of late medieval plague, partial sequencing of the Yersinia pestis pPCP1 plasmid yielded the observation of a 3-T homopolymeric tract which differed from the 5-T homopolymeric tract of the Orientalis Y. pestis CO92 type strain (1). This observation was unexpected because previous data from multispacer sequence typing and glp D gene sequencing yielded only the Orientalis biotype in cases of ancient plague (2).

Using suicide PCR (3), we therefore further investigated pPCP1 in 10 negative control dental pulp specimens and 60 specimens collected from 1 Justinian Orientalis plague site (2), 2 Black Death Orientalis sites, and 2 additional medieval plague sites. All negative controls remained negative; 14 (23%) of 60 plague specimens yielded a PCR product, and 7 interpretable sequences yielded a 3-T homopolymeric tract in all cases.

We further tested a Y. pestis isolate collection comprising 2 Antiqua, 6 Medievalis, and 4 Orientalis strains. No amplification was obtained in DNA-free PCR mix and 5 Y. enterocolitica–negative control isolates, whereas sequencing yielded a 3-T homopolymeric tract in all 12 Y. pestis isolates.

BLAST analysis (http://blast.ncbi.nlm.nih.gov/blast.cgi) indicated that the 5-T homopolymeric tract has been found only once in the Y. pestis CO92 strain (4) and in none of 22 modern and 11 ancient sequences (Table). This 5-T homopolymeric tract is therefore CO92 strain specific and not a marker for the Orientalis biotype. This pPCP1 plasmid sequence, located into a noncoding region of the 3′ extremity of the plasmid, is characterized by several homopolymeric tracts of poly (A) and poly (T), including the 1 herein investigated. Instability of the T-stretches has been reported in bacterial genomes (5) as being hot spots for mutations (5).

Alignment of pPCP1 <italic>Yersinis pestis</italic> modern and ancient sequences
Source and Y. pestis strainGenBank
accession no.Sequence, 5′ → 3′
Complete sequence
Y. pestis CO92 plasmid pPCP1AL109969.18488_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTTTTGTACGCACCACTGAA_8547
Y. pestis KIM plasmid pPCP1AF053945.18488_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_8547
Y. pestis biovar Microtus str. 91001 plasmid pPCP1AE017046.18487_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_8546
Y. pestis Nepal516 plasmid pPCPCP000307.19650_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_9709
Y. pestis Antiqua plasmid pPCPCP000310.19661_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_9720
Y. pestis D182038 plasmid pPCP1CP001592.18486_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_8545
Y. pestis Z176003 plasmid pPCP1
CP001596.1
8487_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_8546
Modern isolate
103813 Y. pestis Nairobi rattus AntiquaHQ54286361_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
103814 Y. pestis JHUPRI AntiquaHQ54286461_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
103815 Y. pestis 14–47 MedievalisHQ54286561_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
103817 Y. pestis 5G5 MedievalisHQ54286661_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
103818 Y. pestis 5F1 MedievalisHQ54286761_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
103819 Y. pestis 6B4 MedievalisHQ54286861_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
103820 Y. pestis 8B7 MedievalisHQ54286961_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
103821 Y. pestis 9F11 MedievalisHQ54287061_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
103822 Y. pestis 6/69M OrientalisHQ54287161_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
103823 Y. pestis EV-76 OrientalisHQ54287261_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCAC_114
103824 Y. pestis algeria 1 OrientalisHQ54287364_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAA_123
103825 Y. pestis algeria 2 Orientalis
HQ542874
61_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
Ancient strain detected from teeth
Tooth no. 107 (excavated from Lariey site, France,
17th century)HQ54287562_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAATGC_123
Tooth no. 515 (excavated from Venice site, Italy,
14th–16th centuries)HQ54287660_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAATGC_121
Tooth no.1183 (excavated from Bondy site, France,
11th–15th centuries)HQ54287762_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_121
Tooth no. 1184 (excavated from Bondy site, France,
11th–15th centuries)HQ54287861_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_121
Tooth no. 1190 (excavated from Bondy site, France,
11th–15th centuries)HQ54287961_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
Tooth no. 254 (excavated from Venice site, Italy,
14th–16th centuries)HQ54288061_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120
Tooth no. 1180 (excavated from Bondy site, France,
11th–15th centuries)HQ54288161_TATATTTTCAAGAAAAGCTGGCTATTTAACATAACGGCAATTT..GTACGCACCACTGAAAT_120

Therefore, in our assessment, the data reported for the late medieval Bavaria burial (1) do not support that deaths of persons buried in this site resulted from a non-Orientalis plague. Typing modern or ancient Y. pestis strains should not rely on poly (A) and poly (T) homopolymeric tracts sequencing.

Suggested citation for this article: Tran T-N-N, Raoult D, Drancourt M. Yersinia pestis DNA sequences in late medieval skeletal finds, Bavaria [letter]. Emerg Infect Dis [serial on the Internet]. 2011 May [date cited]. http://dx.doi.org/10.3201/eid1705.101777

This study was funded by Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes, Unité Mixte de Recherche, Centre National de la Recherche Scientifique 6236.

ReferencesWiechmann I, Harbeck M, Grupe G. Yersinia pestis DNA sequences in late medieval skeletal finds, Bavaria. Emerg Infect Dis. 2010;16:1806–721029555Drancourt M, Roux V, Dang LV, Tran-Hung L, Castex D, Chenal-Francisque V, Genotyping, Orientalis-like Yersinia pestis, and plague pandemics. Emerg Infect Dis. 2004;10:1585–9215498160Raoult D, Aboudharam G, Crubézy E, Larrouy G, Ludes B, Drancourt M. Molecular identification by “suicide PCR” of Yersinia pestis as the agent of medieval Black Death. Proc Natl Acad Sci U S A. 2000;97:12800–3 10.1073/pnas.22022519711058154Parkhill J, Wren BW, Thomson NR, Titball RW, Holden MT, Prentice MB, Genome sequence of Yersinia pestis, the causative agent of plague. Nature. 2001;413:523–7 10.1038/3509708311586360Bechah Y, El Karkouri K, Mediannikov O, Leroy Q, Pelletier N, Robert C, Genomic, proteomic, and transcriptomic analysis of virulent and avirulent Rickettsia prowazekii reveals its adaptive mutation capabilities. Genome Res. 2010;20:655–63 10.1101/gr.103564.10920368341Emerg Infect DisEmerging Infect. DisEIDEmerging Infectious Diseases1080-60401080-6059Centers for Disease Control and Prevention10.3201/eid1705.102013ArticleYersinia pestis DNA Sequences in Late Medieval Skeletal Finds, BavariaWiechmannIngridHarbeckMichaelaGrupeGiselaAuthor affiliations: Ludwig Maximilian University of Munich, Munich, Germany (I. Wiechmann, G. Grupe) Bavarian State Collection of Anthropology and Palaeoanatomy, Munich (M. Harbeck, G. Grupe)Address for correspondence: Ingrid Wiechmann, Ludwig Maximilian University of Munich, Department Biology I, Biodiversity research/Anthropology, Grosshaderner Str. 2, 82152 Planegg-Martinsried, Germany; email: i.wiechmann@lrz.uni-muenchen.deWiechmannI , HarbeckM , GrupeG . Yersinia pestis DNA sequences in late medieval skeletal finds, Bavaria [letter]. Emerg Infect Dis. 2010;17:1806–7.

In Response: We thank Tran et al. for their interest in our article (1). In it, we described the detection of several Yersinia pestis–specific plasmid pPCP1 DNA sequences in skeletal remains from a late medieval mass burial in Bavaria, Germany. In 1 of these sequence sections, we found a deviation from the reference sequence used (Y. pestis strain CO92 plasmid sequence AL109969.1). We did not further interpret this result because we agree with Tran et al. that typing of Y. pestis strains should not rely on poly (A) and poly (T) homopolymeric tract sequencing (2). As we have stated (1), further analyses of our material, including chromosomal markers (3,4) will be conducted to obtain clues as to the specific Y. pestis strain.

Suggested citation for this article: Wiechmann I, Harbeck M, Grupe G. Yersinia pestis DNA sequences in late medieval skeletal finds, Bavaria [letter]. Emerg Infect Dis [serial on the Internet]. 2011 May [date cited]. http://dx.doi.org/10.3201/eid1705.102013

ReferencesWiechmann I, Harbeck M, Grupe G. Yersinia pestis DNA sequences in late medieval skeletal finds, Bavaria [letter] Emerg Infect Dis. 2010;16:1806–721029555Tran T-N-N, Raoult D, Drancourt D. Yersinia pestis DNA sequences in late medieval skeletal finds, Bavaria [letter] Emerg Infect Dis. 2011;17:955–721529431Morelli G, Song Y, Mazzoni CJ, Eppinger M, Roumagnac P, Wagner DM, Yersinia pestis genome sequencing identifies patterns of global phylogenetic diversity. Nat Genet. 2010;42:1140–3 10.1038/ng.70521037571Haensch S, Bianucci R, Signoli M, Rajerison M, Schultz M, Kacki S, Distinct clones of Yersinia pestis caused the Black Death. PLoS Pathog. 2010;6:e1001134 10.1371/journal.ppat.100113420949072