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See detailFATAL ALVEOLAR ECHINOCOCCOSIS OF THE LUMBAR SPINE
KEUTGENS, Aurore ULg; SIMONI, Paolo ULg; DETREMBLEUR, Nancy ULg et al

in Journal of Clinical Microbiology (2013), 51(2), 688-91

For the last ten years, the southern part of Belgium has been recognized as a low-risk endemic area for alveolar echinococcosis. This infection, caused by Echinococcus multilocularis, usually induces a ... [more ▼]

For the last ten years, the southern part of Belgium has been recognized as a low-risk endemic area for alveolar echinococcosis. This infection, caused by Echinococcus multilocularis, usually induces a severe liver condition, and can sometimes spread to other organs. However, alveolar echinococcosis involving bones has been described only very rarely. Here, a fatal case of spondylodiscitis due to E. multilocularis contracted in southern Belgium is reported. [less ▲]

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See detailCapsular Gene Typing of Streptococcus agalactiae Compared to Serotyping by Latex Agglutination
Yao, Kaihu; Poulsen, Knud; Maione, Domenico et al

in Journal of Clinical Microbiology (2012), 51(2), 503-507

We evaluated three different PCR-based capsular gene typing methods applied to 312 human and bovine Streptococcus agalac- tiae (group B Streptococcus [GBS]) isolates and compared the results to serotyping ... [more ▼]

We evaluated three different PCR-based capsular gene typing methods applied to 312 human and bovine Streptococcus agalac- tiae (group B Streptococcus [GBS]) isolates and compared the results to serotyping results obtained by latex agglutination. Among 281 human isolates 27% could not be typed by latex agglutination. All 312 isolates except 5 could be typed by the three PCR methods combined. Two of these methods were multiplex assays. Among the isolates that were typeable by both latex ag- glutination and capsular gene typing, 94% showed agreement between the two methods. However, each of the PCR methods showed limitations. One of the methods did not include all 10 recognized serotypes, one misidentified eight isolates of serotypes Ib and IV as serotype Ia, and one did not distinguish between serotypes VII and IX. For five isolates that showed aberrant pat- terns in the capsular gene typing, long-range PCR targeting the cps operon disclosed large insertions or deletions affecting the cps gene cluster. A sensitive flow cytometric assay based on serotype-specific antibodies applied to 76 selected isolates that were nontypeable by latex agglutination revealed that approximately one-half of these did express capsular polysaccharide. A proce- dure for convenient and reliable capsular gene typing to be included in epidemiological and surveillance studies of S. agalactiae is proposed. [less ▲]

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See detailDiagnostic strategy for identifying avian pathogenic Escherichia coli based on four patterns of virulence genes
Schouler, C; Schaeffer, B; Brée, A et al

in Journal of Clinical Microbiology (2012), 50(5), 1673-1678

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See detailInternational External Quality Assurance for Laboratory Identification and Typing of Streptococcus agalactiae (group B streptococci)
Afshar, Baharak; Broughton, Karen; Creti, Roberta et al

in Journal of Clinical Microbiology (2011)

We report the results from the first international multicentre external quality assessment (EQA) studies for molecular and serological typing of group B streptococcus (GBS) strains as part of DEVANI ... [more ▼]

We report the results from the first international multicentre external quality assessment (EQA) studies for molecular and serological typing of group B streptococcus (GBS) strains as part of DEVANI (DEsign of a Vaccine Against Neonatal Infections), a pan-European programme. A questionnaire-based surveillance was undertaken amongst eight DEVANI and six non-DEVANI participating laboratories from 13 countries in order to assess their current microbiological procedures for GBS screening, diagnosis and typing. GBS strains from three EQA distributions were characterised using molecular and serological methods based on GBS capsular polysaccharide typing. Participants were asked to test the first distribution using their current serotyping and genotyping methods. Strep-B-Latex agglutination method was the most widely used method with a typeability value of >90%. A multiplex PCR assay for GBS capsular gene typing was also used by two of 14 centres who achieved a typeability value of 93%; this assay only detected 9 of 10 GBS capsular polysaccharide genes. From the second and third EQA studies standardised protocols were prepared for serological and molecular typing of GBS strains based on the Strep-B-Latex agglutination method and a novel multiplex PCR assay which detected all ten GBS capsular types (Ia to IX). These standardised protocols are being used by many European laboratories and as the use of these methods increase, it is imperative to continuously improve and assess laboratory performance and offer training to any laboratories that have technical difficulties. [less ▲]

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See detailOriginal findings associated with two cases of bovine papular stomatitis
Dal Pozzo, Fabiana ULg; Martinelle, Ludovic ULg; Gallina, L. et al

in Journal of Clinical Microbiology (2011), 49(12), 4397-4400

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See detailNew assessment of bovine tuberculosis risk factors in Belgium based on nationwide molecular epidemiology.
Humblet, M.-F.; Gilbert, M.; Govaerts, M. et al

in Journal of Clinical Microbiology (2010), 48(8), 2802-8

This assessment aimed to elaborate a statistical nationwide model for analyzing the space-time dynamics of bovine tuberculosis in search of potential risk factors that could be used to better target ... [more ▼]

This assessment aimed to elaborate a statistical nationwide model for analyzing the space-time dynamics of bovine tuberculosis in search of potential risk factors that could be used to better target surveillance measures. A database comprising Mycobacterium bovis molecular profiles from all isolates obtained from Belgian outbreaks during the 1995-to-2006 period (n = 415) allowed the identification of a predominant spoligotype (SB0162). Various databases compiling 49 parameters to be tested were queried using a multiple stepwise logistic regression to assess bovine tuberculosis risk factors. Two isolate datasets were analyzed: the first included all Mycobacterium bovis isolates, while the second included only data related to the SB0162 type strain. When all Mycobacterium bovis isolates were included in the model, several risk factors were identified: history of bovine tuberculosis in the herd (P < 0.001), proximity of an outbreak (P < 0.001), cattle density (P < 0.001), and annual amplitude of mean middle-infrared temperature (P < 0.001). The approach restricted to the predominant SB0162 type strain additionally highlighted the proportion of movements from an infected area during the current year as a main risk factor (P = 0.009). This study identified several risk factors for bovine tuberculosis in cattle, highlighted the usefulness of molecular typing in the study of bovine tuberculosis epidemiology, and suggests a difference of behavior for the predominant type strain. It also emphasizes the role of animals' movements in the transmission of the disease and supports the importance of controlling trade movements. [less ▲]

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See detailAssessment of bovine tuberculosis risk factors based on nationwide molecular epidemiology
Humblet, Marie-France ULg; Gilbert, M.; Govaerts, M. et al

in Journal of Clinical Microbiology (2010)

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See detailCommon virulence factors and genetic relationships between O18 : K1 : H7 Escherichia coli isolates of human and avian origin
Moulin-Schouleur, M.; Schouler, C.; Tailliez, P. et al

in Journal of Clinical Microbiology (2006), 44(10), 3484-3492

Extraintestinal pathogenic (ExPEC) Escherichia coli strains of serotype 018:K1:H7 are mainly responsible for neonatal meningitis and sepsis in humans and belong to a limited number of closely related ... [more ▼]

Extraintestinal pathogenic (ExPEC) Escherichia coli strains of serotype 018:K1:H7 are mainly responsible for neonatal meningitis and sepsis in humans and belong to a limited number of closely related clones. The same serotype is also frequently isolated from the extraintestinal lesions of colibacillosis in poultry, but it is not well known to what extent human and avian strains of this particular serotype are related. Twenty-two ExPEC isolates of human origin and 33 isolates of avian origin were compared on the basis of their virulence determinants, lethality for chicks, pulsed-field gel electrophoresis (PFGE) patterns, and classification in the main phylogenetic groups. Both avian and human isolates were lethal for chicks and harbored similar virulence genotypes. A major virulence pattern, identified in 75% of the isolates, was characterized by the presence of F1 variant fimbriae; S fimbriae; IbeA; the aerobactin system; and genomic fragments A9, A12, D1, D7, D10, and D11 and by the absence of P fimbriae, F1C fimbriae, Afa adhesin, and CNF1. All but one of the avian and human isolates also belonged to major phylogenetic group B2. However, various subclonal populations could be distinguished by PFGE in relation to animal species and geographical origin. These results demonstrate that very closely related clones can be recovered from extraintestinal infections in humans and chickens and suggest that avian pathogenic E. coli isolates of serotype 018:K1:H7 are potential human pathogens. [less ▲]

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See detailExtended-spectrum-beta-lactamase-producing Enterobacteriaceae in Yaounde, Cameroon
Gangoue Pieboji, Joseph ULg; Bedenic, B.; Koulla-Shiro, S. et al

in Journal of Clinical Microbiology (2005), 43(7), 3273-7

Organisms producing extended-spectrum beta-lactamases (ESBLs) have been reported in many countries, but there is no information on the prevalence of ESBL-producing members of the family Enterobacteriaceae ... [more ▼]

Organisms producing extended-spectrum beta-lactamases (ESBLs) have been reported in many countries, but there is no information on the prevalence of ESBL-producing members of the family Enterobacteriaceae in Cameroon. A total of 259 Enterobacteriaceae strains were isolated between 1995 and 1998 from patients at the Yaounde Central Hospital in Cameroon. Enterobacterial isolates resistant to extended-spectrum cephalosporin and monobactam were screened for ESBL production by the double-disk (DD) synergy test. Thirty-one (12%) of these Enterobacteriaceae strains were shown to be positive by the DD synergy test, suggesting the presence of ESBLs. Resistance to oxyimino-cephalosporins and monobactams of 12 (38.7%) of the 31 strains-i.e., 6 Klebsiella pneumoniae, 4 Escherichia coli, 1 Citrobacter freundii, and 1 Enterobacter cloacae strain-was transferred to E. coli HK-225 by conjugation. Resistance to gentamicin, gentamicin plus trimethoprim-sulfamethoxazole, or trimethoprim-sulfamethoxazole was cotransferred into 6, 2, and 1 of these transconjugants, respectively. All 12 transconjugants were resistant to amoxicillin, piperacillin, all of the cephalosporins, and aztreonam but remained susceptible to cefoxitin and imipenem. Crude extracts of beta-lactamase-producing transconjugants were able to reduce the diameters of inhibition zones around disks containing penicillins, narrow- to expanded-spectrum cephalosporins or monobactams when tested against a fully susceptible E. coli strain but had no effect on such zones around cefoxitin, imipenem, and amoxicillin-clavulanate disks. The beta-lactamases produced by the 12 tranconjugants turned out to be SHV-12 by DNA sequencing. Therefore, the ESBL SHV-12 is described for the first time in Cameroon. [less ▲]

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See detailPopulation-level retrospective study of neurologically expressed disorders in ruminants before the onset of bovine spongiform encephalopathy (BSE) in Belgium, a BSE risk III country.
Saegerman, Claude ULg; Berkvens, D.; Claes, L. et al

in Journal of Clinical Microbiology (2005), 43(2), 862-9

A retrospective epidemiological study (n = 7,875) of neurologically expressed disorders (NED) in ruminants before the onset of the bovine spongiform encephalopathy epidemic (years studied, 1980 to 1997 ... [more ▼]

A retrospective epidemiological study (n = 7,875) of neurologically expressed disorders (NED) in ruminants before the onset of the bovine spongiform encephalopathy epidemic (years studied, 1980 to 1997) was carried out in Belgium. The archives of all veterinary laboratories and rabies and transmissible spongiform encephalopathy (TSE) epidemiosurveillance networks were consulted. For all species, a significantly higher number of NED with virological causes (rabies) was reported south of the Sambre-Meuse Valley. During the period 1992 to 1997, for which the data were complete, (i) the predicted annual incidence of NED varied significantly as a function of species and area (higher numbers in areas where rabies was present) but was always above 100 cases per million, and (ii) the mean incidence of suspected TSE cases and, among them, those investigated by histopathological examination varied significantly as a function of species and area. The positive predictive value of a presumptive clinical diagnosis of NED ranged from 0.13 (game) to 0.63 (sheep). Knowledge of the positive predictive value permits the definition of a reference point before certain actions (e.g., awareness and training campaigns) are undertaken. It also shows the usefulness of a systematic necropsy or complementary laboratory tests to establish an etiological diagnosis. TSE analysis of a small, targeted historical sampling (n = 48) permitted the confirmation of one case and uncovered another case of scrapie. The results of the present study help to develop and maintain the quality of the worldwide clinical epidemiological networks for TSE, especially in countries that in the past imported live animals, animal products, and feedstuffs from countries with TSE cases. [less ▲]

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See detailIsolation of shiga toxin-producing Escherichia coli from a South American camelid (Lama guanicoe) with diarrhea.
Mercado, E. C.; Rodriguez, Sabrina ULg; Elizondo, A. M. et al

in Journal of Clinical Microbiology (2004), 42(10), 4809-11

Shiga toxin-producing Escherichia coli belonging to serotype O26:H11 was isolated from a 2-month-old guanaco with severe watery diarrhea. E. coli colonies carried the stx1 and eae genes, showed localized ... [more ▼]

Shiga toxin-producing Escherichia coli belonging to serotype O26:H11 was isolated from a 2-month-old guanaco with severe watery diarrhea. E. coli colonies carried the stx1 and eae genes, showed localized adherence to HEp-2 cells, and produced enterohemolysin. A serological response to lipopolysaccharide O26 was observed at the onset of diarrhea. [less ▲]

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See detailImproved antigenic methods for differential diagnosis of bovine, caprine, and cervine alphaherpesviruses related to bovine herpesvirus 1
Keuser, V.; Schynts, F.; Detry, Bruno et al

in Journal of Clinical Microbiology (2004), 42(3), 1228-1235

The control of infectious bovine rhinotracheitis induced by bovine herpesvirus 1 (BoHV-1) requires sensitive and specific diagnostic assays. As BoHV-1 is antigenically and genetically related to four ... [more ▼]

The control of infectious bovine rhinotracheitis induced by bovine herpesvirus 1 (BoHV-1) requires sensitive and specific diagnostic assays. As BoHV-1 is antigenically and genetically related to four other alphaherpesviruses of ruminants-namely, BoHV-5, caprine herpesvirus 1 (CpHV-1), cervine herpesvirus 1 (CvHV-1) and CvHV-2-diagnostic tests able to discriminate BoHV-1 from these related viruses are needed to avoid misdiagnosis, especially because some of these viruses are able to cross the species barrier. In this study, murine monoclonal antibodies (MAbs) specific for BoHV-1, BoHV-5, CpHV-1, CvHV-1, and CvHV-2 were produced with the aim of setting up an immunofluorescence assay able to discriminate between these related herpesviruses. Produced MAbs were selected for their viral specificity by enzyme-linked immunosorbent assay and indirect immunofluorescence staining of virus-infected cells. Radioimmunoprecipitation characterization of the selected MAbs revealed that four of them are directed against glycoprotein C (gC) and one of them is directed against gD of these related viruses. The obtained results demonstrate that the antibodies produced allow an unambiguous discrimination of each of the four alphaherpesviruses related to BoHV-1. [less ▲]

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See detailDecision support tools for clinical diagnosis of disease in cows with suspected bovine spongiform encephalopathy.
Saegerman, Claude ULg; Speybroeck, N.; Roels, S. et al

in Journal of Clinical Microbiology (2004), 42(1), 172-8

Reporting of clinically suspected cattle is currently the most common method for detecting cases of bovine spongiform encephalopathy (BSE). Improvement of clinical diagnosis and decision-making remains ... [more ▼]

Reporting of clinically suspected cattle is currently the most common method for detecting cases of bovine spongiform encephalopathy (BSE). Improvement of clinical diagnosis and decision-making remains crucial. A comparison of clinical patterns, consisting of 25 signs, was made between all 30 BSE cases, confirmed in Belgium before October 2002, and 272 suspected cases that were subsequently determined to be histologically, immunohistochemically, and scrapie-associated-fiber negative. Seasonality in reporting suspected cases was observed, with more cases being reported during wintertime when animals were kept indoors. The median duration of illness was 30 days. The 10 most relevant signs of BSE were kicking in the milking parlor, hypersensitivity to touch and/or sound, head shyness, panic-stricken response, reluctance to enter in the milking parlor, abnormal ear movement or carriage, increased alertness behavior, reduced milk yield, teeth grinding, and temperament change. Ataxia did not appear to be a specific sign of BSE. A classification and regression tree was constructed by using the following four features: age of the animal, year of birth, number of relevant BSE signs noted, and number of clinical signs, typical for listeriosis, noted. The model had a sensitivity of 100% and a specificity of 85%. This approach allows the use of an interactive decision-support tool, based entirely on odds ratios, a statistic independent of disease prevalence. [less ▲]

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See detailMultiplex PCRs for identification of necrotoxigenic Escherichia coli
Van Bost, S.; Jacquemin, E.; Oswald, E. et al

in Journal of Clinical Microbiology (2003), 41(9), 4480-4482

Two multiplex PCRs were developed for the detection of necrotoxigenic Escherichia coli virulence genes. M1 contained the primers for the toxins and the aerobactin, and M2 contained the primers for the ... [more ▼]

Two multiplex PCRs were developed for the detection of necrotoxigenic Escherichia coli virulence genes. M1 contained the primers for the toxins and the aerobactin, and M2 contained the primers for the adhesins. They were validated by single PCRs performed with reference E. coli strains and by multiplex PCRs with necrotoxigenic E. coli strains isolated from different animal species. [less ▲]

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See detailDeep infection by Trichophyton rubrum in an immunocompromised patient.
Nir-Paz, Ran; Elinav, Hila; Pierard, Gérald ULg et al

in Journal of Clinical Microbiology (2003), 41(11), 5298-301

Dermatophytes are common pathogens of skin but rarely cause invasive disease. We present a case of deep infection by Trichophyton rubrum in an immunocompromised patient. T. rubrum was identified by ... [more ▼]

Dermatophytes are common pathogens of skin but rarely cause invasive disease. We present a case of deep infection by Trichophyton rubrum in an immunocompromised patient. T. rubrum was identified by morphological characteristics and confirmed by PCR. Invasiveness was apparent by histopathology and immunohistochemistry. The patient was treated successfully with itraconazole. [less ▲]

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See detailDetection of Aspergillus spp. by PCR in Bronchoalveolar Lavage Fluid
Hayette, Marie-Pierre ULg; Vaira, Dolorès ULg; Suzin, Fabrice et al

in Journal of Clinical Microbiology (2001), 39(6), 2338-2340

The usefulness of a nested PCR assay for detection of Aspergillus sp. DNA was evaluated in 177 bronchoalveolar lavage (BAL) fluid specimens. This test was accurate both to diagnose culture-negative BAL ... [more ▼]

The usefulness of a nested PCR assay for detection of Aspergillus sp. DNA was evaluated in 177 bronchoalveolar lavage (BAL) fluid specimens. This test was accurate both to diagnose culture-negative BAL fluid specimens from patients with invasive pulmonary aspergillosis and to confirm culture-positive samples. However, it did not differentiate between infection and colonization. [less ▲]

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See detailProduction of Bovine Herpesvirus Type 1-Seronegative Latent Carriers by Administration of a Live-Attenuated Vaccine in Passively Immunized Calves
Lemaire, Mylène; Meyer, Gilles; Baranowski, Eric et al

in Journal of Clinical Microbiology (2000), 38(11), 4233-8

The consequences of the vaccination of neonatal calves with the widely used live-attenuated temperature-sensitive (ts) bovine herpesvirus type 1 (BHV-1) were investigated. The ts strain established acute ... [more ▼]

The consequences of the vaccination of neonatal calves with the widely used live-attenuated temperature-sensitive (ts) bovine herpesvirus type 1 (BHV-1) were investigated. The ts strain established acute and latent infections in all vaccinated calves either with or without passive immunity. Four of seven calves vaccinated under passive immunity became clearly BHV-1 seronegative by different serological tests, as did uninfected control calves after the disappearance of maternal antibodies, and they remained so for long periods. A cell-mediated immune response was detected by a BHV-1 gamma interferon assay, but this test failed to detect the seronegative latent carriers (SNLCs). While they are not detected, SNLCs represent a threat for BHV-1-free herds or countries. This study demonstrates that SNLCs can be easily obtained by inoculation with a live-attenuated BHV-1 under passive immunity and that latent carrier animals without any antibody do exist. Consequently, this situation could represent a good model to experimentally produce SNLCs. [less ▲]

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See detailEffects of Bovine Herpesvirus Type 1 Infection in Calves with Maternal Antibodies on Immune Response and Virus Latency
Lemaire, Mylène; Weynants, Vincent; Godfroid, Jacques et al

in Journal of Clinical Microbiology (2000), 38(5), 1885-94

The presence of maternally derived antibodies can interfere with the development of an active antibody response to antigen. Infection of seven passively immunized young calves with a virulent strain of ... [more ▼]

The presence of maternally derived antibodies can interfere with the development of an active antibody response to antigen. Infection of seven passively immunized young calves with a virulent strain of bovine herpesvirus type 1 (BHV-1) was performed to determine whether they could become seronegative after the disappearance of maternal antibodies while latently infected with BHV-1. Four uninfected calves were controls. All calves were monitored serologically for 13 to 18 months. In addition, the development of a cell-mediated immune response was assessed by an in vitro antigen-specific gamma interferon (IFN-gamma) production assay. All calves had positive IFN-gamma responses as early as 7 days until at least 10 weeks after infection. However, no antibody rise was observed after infection in the three calves with the highest titers of maternal antibodies. One of the three became seronegative by virus neutralization test at 7 months of age like the control animals. This calf presented negative IFN-gamma results at the same time and was classified seronegative by enzyme-linked immunosorbent assay at around 10 months of age. This calf was latently infected, as proven by virus reexcretion after dexamethasone treatment at the end of the experiment. In conclusion, this study demonstrated that BHV-1-seronegative latent carriers can be obtained experimentally. In addition, the IFN-gamma assay was able to discriminate calves possessing only passively acquired antibodies from those latently infected by BHV-1, but it could not detect seronegative latent carriers. The failure to easily detect such animals presents an epidemiological threat for the control of BHV-1 infection. [less ▲]

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