@article {47227, title = {Detection and culture of Bartonella quintana, Serratia marcescens, and Acinetobacter spp. from decontaminated human body lice}, journal = {Journal of Clinical Microbiology}, volume = {39}, year = {2001}, month = {2001}, pages = {1707 - 1709}, url = {http://www.phthiraptera.info/sites/phthiraptera.info/files/46338.pdf}, author = {La Scola,Bernard and Pierre-Edouard Fournier and Brouqui,Philippe and Didier Raoult} } @article {46237, title = {Body lice as tools for diagnosis and surveillance of reemerging diseases}, journal = {Journal of clinical microbiology}, volume = {37}, year = {1999}, month = {1999}, pages = {596 - 599}, abstract = {Body lice are vectors of three bacteria which cause human disease: Rickettsia prowazekii, the agent of epidemic typhus; Bartonella quintana, the agent of trench fever; and Borrelia recurrentis, the agent of relapsing fever. A recrudescence of body lice is being observed as the numbers of individuals living under social conditions which predispose individuals to infestation have increased. Because this phenomenon may lead to the reemergence of infections transmitted by body lice, we aimed to assess the occurrence and prevalence of the three agents described above in more than 600 body lice collected from infested individuals in the African countries of Congo, Zimbabwe, and Burundi, in France, in Russia, and in Peru. The presence of the three bacteria in each louse was determined by specific PCR amplification, and the identities of the organisms detected were confirmed by determination of the nucleotide base sequences of the amplification products. Using this approach, we were able to confirm the presence of R. prowazekii in lice collected from refugees in Burundi, among whom typhus was epidemic, and the presence of B. quintana in lice collected from all locations except the Congo. B. recurrentis was never found. Molecular approaches are convenient tools for the detection and identification of bacterial DNA in body lice and for the epidemiological study of louse-borne bacteria from countries where no medical and biological laboratory facilities are available.}, keywords = {animals, Bartonella quintana/isolation \& purification, Borrelia, Burundi, Congo, France, homeless, humans, Insect vectors, Lice Infestations/complications/epidemiology, Pediculus, Peru, polymerase, populations, Relapsing Fever, Rickettsia prowazekii/isolation \& purification, Russia/epidemiology, trench fever, typhus, Zimbabwe}, url = {http://phthiraptera.info/sites/phthiraptera.info/files/46237.pdf}, author = {Roux,Veronique and Didier Raoult} } @article {46145, title = {Serodiagnosis of Louse-Borne relapsing fever with glycerophosphodiester phosphodiesterase (GlpQ) from Borrelia recurrentis}, journal = {Journal of clinical microbiology}, volume = {38}, year = {2000}, note = {LR: 20041117; JID: 7505564; 0 (Bacterial Proteins); 0 (DNA Primers); 0 (DNA, Bacterial); EC 3.1.4 (Phosphoric Diester Hydrolases); EC 3.1.4.46 (glpQ protein, Bacteria); ppublish}, month = {2000}, pages = {3561 - 3571}, address = {UNITED STATES}, abstract = {

Human louse-borne relapsing fever occurs in sporadic outbreaks in central and eastern Africa that are characterized by significant morbidity and mortality. Isolates of the causative agent, Borrelia recurrentis, were obtained from the blood of four patients during a recent epidemic of the disease in southern Sudan. The glpQ gene, encoding glycerophosphodiester phosphodiesterase, from these isolates was sequenced and compared with the glpQ sequences obtained from other relapsing-fever spirochetes. Previously we showed that GlpQ of Borrelia hermsii is an immunogenic protein with utility as a serological test antigen for discriminating tick-borne relapsing fever from Lyme disease. In the present work, we cloned and expressed the glpQ gene from B. recurrentis and used recombinant GlpQ in serological tests. Acute- and convalescent-phase serum samples obtained from 42 patients with louse-borne relapsing fever were tested with an indirect immunofluorescence assay (IFA) and an enzyme-linked immunosorbent assay (ELISA) that used whole cells of B. recurrentis and with immunoblotting to whole-cell lysates of the spirochete and Escherichia coli producing recombinant GlpQ. The geometric mean titers of the acute- and convalescent-phase serum samples measured by IFA were 1:83 and 1:575, respectively. The immunoblot analysis identified a high level of reactivity and seroconversion to GlpQ, and the assay was more sensitive than the whole-cell IFA and ELISA using purified, recombinant histidine-tagged GlpQ. Serum antibodies to GlpQ and other antigens persisted for 27 years in one patient. We conclude that assessment of anti-GlpQ antibodies will allow serological confirmation of louse-borne relapsing fever and determination of disease prevalence.

}, keywords = {Amino Acids, animals, Bacterial Proteins/analysis/genetics, Borrelia, Cloning, Molecular, dna, Ethiopia, Fluorescent Antibody Technique, Indirect, humans, Insect vectors, Lice/microbiology, Phylogeny, polymerase, sequence, Serologic Tests}, isbn = {0095-1137}, doi = {https://doi.org/10.1128/JCM.38.10.3561-3571.2000}, url = {https://journals.asm.org/doi/full/10.1128/JCM.38.10.3561-3571.2000}, author = {Porcella,S. F. and Raffel,S. J. and Schrumpf,M. E. and Schriefer,M. E. and Dennis,D. T. and Schwan,T. G.} } @article {46093, title = {Detection and culture of Bartonella quintana, Serratia marcescens, and Acinetobacter spp. from decontaminated human body lice}, journal = {Journal of Clinical Microbiology}, volume = {39}, year = {2001}, month = {2001}, pages = {1707 - 1709}, abstract = {As part of a survey for trench fever among homeless people in Marseilles, France, we attempted isolation of Bartonella quintana from body lice. A decontamination protocol of immersion in 70\% ethanol with 0.2\% iodine was devised and was tested with a laboratory colony of body lice. Lice which had been experimentally contaminated with either Escherichia coli, Staphylococcus epidermidis, or Acinetobacter spp. were successfully decontaminated, and this process did not prevent the culture of B. quintana from these lice. One hundred sixty-one lice obtained from homeless patients were studied by the protocol. B. quintana was isolated on axenic medium from 15 of 161 body lice and was detected in 41 of 161 lice by PCR. Acinetobacter spp. and Serratia marcescens were also isolated from body lice. The sensitivities of PCR and culture of B. quintana were 98 and 36\%, respectively. These procedures may be useful for epidemiologic studies of trench fever and for the recovery of strains for characterization and comparison.}, keywords = {Acinetobacter, animals, Bartonella quintana/classification/genetics/isolation \& purification, Culture Media, dna, homeless, humans, Iodine/pharmacology, Pediculus, polymerase, Sensitivity and Specificity, Serratia, trench fever}, url = {http://www.phthiraptera.info/sites/phthiraptera.info/files/44916.pdf}, author = {La Scola,Bernard and Pierre-Edouard Fournier and Brouqui,Philippe and Didier Raoult} } @article {46043, title = {Detection of Rickettsia prowazekii in body lice and their feces by using monoclonal antibodies}, journal = {Journal of Clinical Microbiology}, volume = {40}, year = {2002}, month = {2002}, pages = {3358 - 3363}, abstract = {

In order to identify Rickettsia prowazekii in lice, we developed a panel of 29 representative monoclonal antibodies selected from 187 positive hybridomas made by fusing splenocytes of immunized mice with SP2/0-Ag14 myeloma cells. Immunoblotting revealed that 15 monoclonal antibodies reacted with the lipopolysaccharide-like (LPS-L) antigen and 14 reacted with the epitopes of a 120-kDa protein. Only typhus group rickettsiae reacted with the monoclonal antibodies against LPS-L. R. felis, a recently identified rickettsial species, did not react with these monoclonal antibodies, confirming that it is not antigenically related to the typhus group. Monoclonal antibodies against the 120-kDa protein were highly specific for R. prowazekii. We successfully applied a selected monoclonal antibody against the 120-kDa protein to detect by immunofluorescence assay R. prowazekii in smears from 56 wild and laboratory lice, as well as in 10 samples of louse feces infected or not infected with the organism. We have developed a simple, practical, and specific diagnostic assay for clinical specimens and large-scale epidemiological surveys with a sensitivity of 91\%. These monoclonal antibodies could be added to the rickettsial diagnostic panel and be used to differentiate R. prowazekii from other rickettsial species.

}, keywords = {animals, Antibodies, Bacterial/biosynthesis/diagnostic use/immunology, Antibodies, Monoclonal/biosynthesis/diagnostic use/immunology, Antibody Specificity, Blotting, Western, electrophoresis, Feces/microbiology, mice, Mice, Inbred BALB C, Pediculus, Rickettsia prowazekii/immunology/isolation \& purification}, url = {http://www.phthiraptera.info/sites/phthiraptera.info/files/44853.pdf}, author = {Fang,Rong and Houhamdi,Linda and Didier Raoult} } @article {45898, title = {Multispacer typing technique for sequence-based typing of Bartonella quintana}, journal = {Journal of clinical microbiology}, volume = {43}, year = {2005}, note = {GENBANK/AY660702; GENBANK/AY660703; GENBANK/AY660704; GENBANK/AY660705; GENBANK/AY660706; GENBANK/AY660707; GENBANK/AY660708; GENBANK/AY660709; GENBANK/AY660710; GENBANK/AY660711; GENBANK/AY660712; GENBANK/AY660713; GENBANK/AY660714; GENBANK/AY660715; GENBANK/AY660716; GENBANK/AY660717; GENBANK/AY660718; JID: 7505564; 0 (DNA, Bacterial); 0 (DNA, Ribosomal Spacer); ppublish}, month = {2005}, pages = {41 - 48}, abstract = {Bartonella quintana is a worldwide fastidious bacterium of the Alphaproteobacteria responsible for bacillary angiomatosis, trench fever, chronic lymphadenopathy, and culture-negative endocarditis. The recent genome sequencing of a B. quintana isolate allowed us to propose a genome-wide sequence-based typing method. To ensure sequence discrimination based on highly polymorphic areas, we amplified and sequenced 34 spacers in a large collection of B. quintana isolates. Six of these exhibited polymorphisms and allowed the characterization of 4 genotypes. However, the strain variants suggested by the noncoding sequences did not correlate with the results of pulsed-field gel electrophoresis (PFGE), which suggested a higher degree of variability. Modification of the PFGE profile of one isolate after nine subcultures confirmed that rearrangement frequencies are high in this species, making PFGE unreliable for epidemiological purposes. The low extent of sequence heterogeneity in the species suggests a recent emergence of this bacterium as a human pathogen. Direct typing of natural samples allowed the identification of a fifth genotype in the DNA extracted from a human body louse collected in Burundi. We have named the typing technique herein described multispacer typing.}, keywords = {animals, Bacteremia/microbiology, Bacterial Typing Techniques, Bartonella quintana/classification/genetics/isolation \& purification, dna, electrophoresis, Genotype, humans, Lice/microbiology, Molecular Sequence Data, Research Support, Non-U.S. Gov{\textquoteright}t, sequence, trench fever}, isbn = {0095-1137}, author = {Foucault,C. and La Scola,B. and Lindroos,H. and Andersson,S. G. and Didier Raoult} } @article {45853, title = {Multispacer typing of Rickettsia prowazekii enabling epidemiological studies of epidemic typhus}, journal = {Journal of Clinical Microbiology}, volume = {43}, year = {2005}, note = {GENBANK/AY695447; GENBANK/AY695448; GENBANK/AY695449; GENBANK/AY695452; GENBANK/AY695454; JID: 7505564; 0 (Bacterial Proteins); 0 (DNA, Bacterial); 0 (DNA, Ribosomal Spacer); ppublish}, month = {2005}, pages = {4708 - 4712}, abstract = {

Currently, there is no tool for typing Rickettsia prowazekii, the causative agent of epidemic typhus, currently considered a potential bioterrorism agent, at the strain level. To test if the multispacer typing (MST) method could differentiate strains of R. prowazekii, we amplified and sequenced the 25 most variable intergenic spacers between the R. prowazekii and R. conorii genomes in five strains and 10 body louse amplicons of R. prowazekii from various geographic origins. Two intergenic spacers, i.e., rpmE/tRNA(fMet) and serS/virB4, were variable among tested R. prowazekii isolates and allowed identification of three and two genotypes, respectively. When the genotypes obtained from the two spacers were combined, we identified four different genotypes. MST demonstrated that several R. prowazekii strains circulated in human body lice during an outbreak of epidemic typhus in Burundi. This may help to discriminate between natural and intentional outbreaks. Our study supports the usefulness of MST as a versatile method for rickettsial strain genotyping.

}, keywords = {animals, Bacterial Proteins, Bacterial Typing Techniques, Disease Outbreaks, dna, humans, Lice/microbiology, Molecular Sequence Data, polymerase, Rickettsia prowazekii/classification/genetics/isolation \& purification, sequence, typhus, variation}, isbn = {0095-1137}, author = {Zhu,Y. and Pierre-Edouard Fournier and Ogata,H. and Didier Raoult} } @article {40250, title = {Rickettsia felis: a new species of pathogenic Rickettsia isolated from cat fleas}, journal = {Journal of Clinical Microbiology}, volume = {34}, year = {1996}, month = {1996}, pages = {671 - 674}, abstract = {A flea-borne rickettsia, previously referred to as ELB, has been implicated as a cause of human illness, Using sequence data obtained from a fragment of the citrate synthase gene, we compared ELB, Rickettsia australis, R. rickettsii, and R. akari with the louse-borne R. prowazekii. We tallied 24 base pair differences between ELB and R. prowazekii and 25 between R. rickettsii and R. prowazekii; there were 30 base pair differences between R. australis and R. prowazekii and 29 between R. akari and R. prowazekii. We observed 32 differences between Rickettsia typhi and ELB. Sodium dodecyl sulfate- polyacrylamide gel electrophoresis and immunoblot analyses of ELB, with typing sera against R. typhi indicate that ELB surface antigens are more closely related to the flea-borne R. typhi than to the mite-borne R. akari, On the basis of the results of citrate synthase gene sequence comparisons, as well as previous comparisons with 16S rRNA and 17-kDa-protein gene segments, we found that ELB is sufficiently genetically distinct from other rickettsiae to be designated a new species, Rickettsia felis.}, keywords = {typhus}, author = {Higgins,J. A. and Radulovic,S. and Schriefer,M. E. and Azad,A. F.} }