B. Jaulhac

RELEVANCE OF THE ANTIBODY INDEX TO DIAGNOSE LYME NEUROBORRELIOSIS AMONG SEROPOSITIVE PATIENTS

Background: No consensual criteria exist to diagnose neuroborreliosis. The intrathecal anti-Borrelia antibody index (AI) is a necessary criterion to diagnose neuroborreliosis in Europe, but not in the United States. Previous studies to determine the diagnostic value of the AI found a sensitivity ranging from 55% to 80%. However, these studies included only typical clinical cases of meningitis or meningoradiculitis, and none had a control group with CSF anti-Borrelia antibodies. Methods: We studied a sample of 123 consecutive patients with clinical signs of neurologic involvement and CSF anti-Borrelia antibodies. We determined the AI for all patients and a final diagnosis was made. Patients were then divided into three groups (neuroborreliosis, possible neuroborreliosis, control). Results: Thirty of the 40 patients with neuroborreliosis had a positive AI (AI sensitivity = 75%). Two of the 74 patients with another neurologic diagnosis had a positive AI (AI specificity = 97%). Conclusion: The antibody index has a very good specificity but only moderate sensitivity. Given the lack of consensual criteria for neuroborreliosis and the absence of a “gold standard” diagnostic test, we propose pragmatic diagnostic criteria for neuroborreliosis, namely the presence of four of the following five items: no past history of neuroborreliosis, positive CSF ELISA serology, positive anti-Borrelia antibody index, favorable outcome after specific antibiotic treatment, and no differential diagnosis. These new criteria will need to be tested in a larger, prospective cohort.

Diagnosis of Cat Scratch Disease with Detection of Bartonella henselae by PCR: a Study of Patients with Lymph Node Enlargement

ABSTRACT Cat scratch disease (CSD) is mostly due to Bartonella henselae after inoculation of the organism through a skin injury. Since the causative bacteria cannot be easily cultured from human lymph node samples, the diagnosis usually relies on epidemiological, clinical, histological, and serological criteria (classical criteria). A study was performed to determine the diagnostic value of PCR analysis for the detection of B. henselae for the diagnosis of CSD and its place in the diagnostic strategy alongside the classical criteria. Over a 7-year period, lymph node biopsy specimens or cytopunctures from 70 patients were systematically tested by PCR for the presence of B. henselae DNA (htrA gene) in the Bacteriology Laboratory of the Hôpitaux Universitaires de Strasbourg. Serological testing by an immunofluorescence assay for B. henselae antibodies was also performed for each patient, and clinical, epidemiological, and histological data were collected. The patients were then divided into two groups according to the number of positive diagnostic criteria for CSD: 29 patients with definite CSD (two or more classical criteria) and 15 patients with possible CSD (less than two classical criteria). The remaining 26 patients for whom another diagnosis was retained were used as a control group. Among all criteria, PCR analysis had the best specificity (100%). The PCR assay for B. henselae was positive for 22 (76%; 95% confidence interval [CI95], 56.5 to 89.7%) of the 29 definite CSD patients and 3 (20%; CI95, 4.3 to 48.1%) of the 15 possible CSD patients. We then studied combinations of diagnostic criteria, including B. henselae PCR analysis. The best diagnostic performance was observed if at least two criteria were present among serologic, epidemiologic, histological, and molecular criteria.

Dialister pneumosintes Associated with Human Brain Abscesses

ABSTRACT In this report, we review two cases of brain infection due to Dialister pneumosintes in previously healthy patients. The bacterium was isolated from the first patient by blood culture and directly from a brain abscess in the second patient. In both cases, the infection was suspected to be of nasopharyngeal or dental origin. The patients had favorable outcomes following surgical debridement and antibiotic treatment. After in vitro amplification and partial sequencing of the 16S rRNA gene, two strains were classified as D. pneumosintes. However, traditional biochemical tests were not sufficient to identify the bacteria. In addition to causing periodontal and opportunistic infections, D. pneumosintes, contained in mixed flora, may behave as a clinically important pathogen, especially in the brain. In addition to phenotypic characterization, 16S rRNA partial sequencing was used to identify D. pneumosintes definitively.

Peripartum Bacteremias Due to Leptotrichia amnionii and Sneathia sanguinegens, Rare Causes of Fever during and after Delivery

ABSTRACT We report three cases of delivery and postpartum bacteremia due to unusual anaerobic bacteria in healthy young women. Leptotrichia amnionii bacteremia occurred during delivery in two mothers and was associated with fetal distress during labor. Conversely, Sneathia sanguinegens bacteremia occurred postpartum, 2 days after delivery, without consequence for the neonate.

Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry-Based Single Nucleotide Polymorphism Genotyping Assay Using iPLEX Gold Technology for Identification of Mycobacterium tuberculosis Complex Species and Lineages

ABSTRACT The major goal of the present study was to investigate the potential use of a novel single nucleotide polymorphism (SNP) genotyping technology, called iPLEX Gold (Sequenom), for the simultaneous analysis of 16 SNPs that have been previously validated as useful for identification of Mycobacterium tuberculosis complex (MTBC) species and classification of MTBC isolates into distinct genetic lineages, known as principal genetic groups (PGGs) and SNP cluster groups (SCGs). In this context, we developed a 16-plex iPLEX assay based on an allele-specific-primer single-base-extension reaction using the iPLEX Gold kit (Sequenom), followed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis on the commercially available Sequenom MassARRAY platform. This assay was tested on a panel of 55 well-characterized MTBC strains that were also genotyped for the same loci using the previously reported SNaPshot assay, as well as 10 non-MTBC mycobacteria and 4 bacteria not belonging to the genus Mycobacterium. All MTBC samples were successfully analyzed with the iPLEX assay, which yielded clear allelic data for 99.9% of the SNPs (879 out of 880). No false-positive results were obtained with the negative controls. Compared to the SNaPshot assay, the newly developed 16-plex iPLEX assay produced fully concordant results that allowed reliable differentiation of MTBC species and recognition of lineages, thus demonstrating its potential value in diagnostic, epidemiological, and evolutionary applications. Compared to the SNaPshot approach, the implementation of the iPLEX technology could offer a higher throughput and could be a more flexible and cost-effective option for microbiology laboratories.

Microarray Analyses of Inflammation Response of Human Dermal Fibroblasts to Different Strains of Borrelia burgdorferi Sensu Stricto

In Lyme borreliosis, the skin is the key site of bacterial inoculation by the infected tick, and of cutaneous manifestations, erythema migrans and acrodermatitis chronica atrophicans. We explored the role of fibroblasts, the resident cells of the dermis, in the development of the disease. Using microarray experiments, we compared the inflammation of fibroblasts induced by three strains of Borrelia burgdorferi sensu stricto isolated from different environments and stages of Lyme disease: N40 (tick), Pbre (erythema migrans) and 1408 (acrodermatitis chronica atrophicans). The three strains exhibited a similar profile of inflammation with strong induction of chemokines (CXCL1 and IL-8) and IL-6 cytokine mainly involved in the chemoattraction of immune cells. Molecules such as TNF-alpha and NF-κB factors, metalloproteinases (MMP-1, -3 and -12) and superoxide dismutase (SOD2), also described in inflammatory and cellular events, were up-regulated. In addition, we showed that tick salivary gland extracts induce a cytotoxic effect on fibroblasts and that OspC, essential in the transmission of Borrelia to the vertebrate host, was not responsible for the secretion of inflammatory molecules by fibroblasts. Tick saliva components could facilitate the early transmission of the disease to the site of injury creating a feeding pit. Later in the development of the disease, Borrelia would intensively multiply in the skin and further disseminate to distant organs.

Species of Borrelia burgdorferi complex that cause borrelial lymphocytoma in France

Backgroundu2002 Only about 30 cases of borrelial lymphocytoma (BL) with identification of the causative species of Borrelia have been published to date, mainly from Eastern or Central European countries.

Myélite aiguë et neuroborréliose

Resume Introduction Les myelites aigues de Lyme representent 4 a 5 p. 100 des cas de neuroborreliose. Seuls 8 cas ayant fait l’objet a la fois d’une etude du liquide cephalo-rachidien et d’une IRM medullaire ont ete decrits dans la litterature. Nous rapportons ici 3 nouveaux cas. Methode Dans une serie de 45 patients atteints de neuroborreliose diagnostiques en 8 ans entre le 1er janvier 1997 et le 31 decembre 2004, 3 avaient un tableau de myelite aigue. Nous en avons analyse les caracteristiques cliniques, biologiques et radiologiques. Observations Ces 3 patients avaient tous une atteinte motrice, sensitive et sphincterienne a des degres divers. Chaque patient avait aussi une atteinte extra-medullaire : fievre et cephalees pour l’un, paralysie faciale peripherique unilaterale pour le second et hemorragie meningee pour le dernier. Il existait une pleiocytose d’intensite variable de 10 a 520 globules blancs par mm3. La serologie de Lyme etait positive dans le liquide cephalo-rachidien dans les 3 cas. L’index de synthese intrathecale anti-Borrelia burgdorferi etait positif ou intermediaire. L’IRM medullaire retrouvait un hypersignal de plus de 3 metameres de haut pouvant etre transverse, posterieur ou centro-medullaire. L’evolution sous antibiotherapie specifique pendant au moins 3 semaines etait favorable pour les 3 cas. Conclusion Ces 3 cas ainsi que les autres decrits dans la litterature montrent la diversite des tableaux clinico-radiologiques de myelite aigue de Lyme : myelite transverse, posterieure ou centrale. Ainsi, une serologie de Lyme dans le liquide cephalo-rachidien doit etre systematiquement pratiquee dans les myelites aigues etendues, en particulier en zone d’endemie.

Proteomic analysis of three Borrelia burgdorferi sensu lato native species and disseminating clones: Relevance for Lyme vaccine design

Lyme borreliosis is the most important vector‐borne disease in the Northern hemisphere. It is caused by Borrelia burgdorferi sensu lato bacteria transmitted to humans by the bite of hard ticks, Ixodes spp. Although antibiotic treatments are efficient in the early stage of the infection, a significant number of patients develop disseminated manifestations (articular, neurological, and cutaneous) due to unnoticed or absence of erythema migrans, or to inappropriate treatment. Vaccine could be an efficient approach to decrease Lyme disease incidence. We have developed a proteomic approach based on a one dimensional gel electrophoresis followed by LC‐MS/MS strategy to identify new vaccine candidates. We analyzed a disseminating clone and the associated wild‐type strain for each major pathogenic Borrelia species: B. burgdorferi sensu stricto, B. garinii, and B. afzelii. We identified specific proteins and common proteins to the disseminating clones of the three main species. In parallel, we used a spectral counting strategy to identify upregulated proteins common to the clones. Finally, 40 proteins were found that could potentially be involved in bacterial virulence and of interest in the development of a new vaccine. We selected the three proteins specifically detected in the disseminating clones of the three Borrelia species and checked by RT‐PCR whether they are expressed in mouse skin upon B. burgdorferi ss inoculation. Interestingly, BB0566 appears as a potential vaccine candidate. All MS data have been deposited in the ProteomeXchange with identifier PXD000876 (http://proteomecentral.proteomexchange.org/dataset/PXD000876).

Manifestations infectieuses systémiques post piqûres de tiques : étude étiologique et place des coinfections

Introduction Les tiques transmettent plusieurs agents pathogenes pour l’homme. L’hypothese de coinfections est avancee pour expliquer des tableaux cliniques complexes. Nous avons realise une etude prospective sur 2xa0ans dans le but d’identifier les causes infectieuses des manifestations aigues post piqures de tiques. Materiels et methodes Critere d’inclusionxa0: patients avec manifestation clinique dans le mois qui a suivi une piqure de tique, ou patient expose aux tiques (frequentation des forets, habitat en milieu rural), presentant une fievre sans autre cause evidente. Realisation systematique de frottis sanguins ( Anaplasma , Babesia ), PCR Anaplasma , serologie a 4xa0a 6xa0semaines d’intervalle ( Borrelia , TBE, Anaplasma , Bartonella , Francisella ). Le medecin etait libre de prescrire d’autres examens selon l’orientation clinique. Criteres diagnostiquesxa0: anaplasmosexa0: PCRxa0+xa0ou serologiexa0>xa01/128xa0ou seroconversionxa0; bartonellosexa0: seroconversion ou ascensionxa0×xa04xa0du taux des Acxa0; tularemiexa0: serologiexa0+xa0CNRxa0; Lymexa0: EM ou seroconversion ou IgM, babesiosexa0: frottis sanguin, TBExa0: IgM et IgG+. Resultats Au total, 129xa0patients repondant aux criteres d’inclusion ayant complete les deux visites ont ete analyses. Pour 55xa0patients un diagnostic a ete etabli. Pour les infections transmises par les tiquesxa0: 19xa0Anaplasmose, 10xa0borreliose de Lyme (7xa0EMxa0; 6xa0patients avec presence d’IgM (dont 2xa0IgM uniquementxa0+xa0a V2xa0; un cas avec IgM isolees n’a pas ete retenu car il presentait une primo infection CMV), 7xa0infections a TBE, 1xa0tularemie. Pour les bartonelloses, 7xa0patients avait une serologiexa0>xa01/64xa0a V2, dont une seule ascension significative chez un patient ayant fait une leptospirose confirmee. Parmi les autres diagnosticsxa0: 6xa0infections a CMV, 4xa0leptospirose, 2xa0infection a EBV, 1xa0infection a Hantavirus , 1xa0paludisme, 1xa0primo infection VIH, 1xa0infection a Parvovirus, 1xa0bacteriemie a Aeromonas . Dans trois cas nous avons pu identifier une possible coinfection. Deux patients avec une association TBE et Lyme, reposant une fois sur l’identification de Borrelia dans le sang (coinfection confirmee) et une fois sur la presence d’IgM Borrelia , sans seroconversion IgG (fauxxa0+xa0possible). Un patient avec une association TBE confirmee, Lyme (IgM Borrelia sans seroconversion IgGxa0: fauxxa0+xa0possible) et anaplasmose (serologie Anaplasma 1/128, avec PCR negativexa0: fauxxa0+xa0possible). Conclusion Les 3xa0diagnostics les plus frequents sont l’anaplasmose, la borreliose de Lyme et les infections a virus TBE. Les coinfections sont rares avec des risques de serologies faussement positives. L’exposition a une piqure de tique ne doit pas faire meconnaitre les autres causes d’infection.