bluuebunny/crossref_metadata_2025_split huggingface dataset:
Multicenter Comparison of BACTEC 9050 and BACTEC 9240 Blood Culture Systems
Authors: Murray Patrick R. , Hollick Gary E. , Jerris Robert C. , Wilson Michael L. 
Published: December 2019

ABSTRACT

The overall recovery of organisms and time to detection with the BACTEC 9050 and BACTEC 9240 systems were compared in a multicenter evaluation. In the first phase of the study, a total of 4,383 compliant aerobic (Plus Aerobic/F) blood culture sets were processed. There was no significant difference in the recovery of individual groups of organisms with the two systems, with the exception of
Streptococcus pneumoniae
which was isolated more frequently with BACTEC 9050. False-positive signals occurred more often with BACTEC 9240 (58 cultures) than with BACTEC 9050 (43 cultures), but false-negative cultures were uncommon with both systems (3 cultures for each system). Time to detection of positive cultures of clinically significant organisms was essentially the same with both instruments. In the second phase of the study, 2,431 compliant anaerobic (Plus Anaerobic/F) blood culture sets were processed. There was no significant difference in the recovery of organisms with BACTEC 9050 compared with BACTEC 9240. Significantly (
P
< 0.03) more false-positive signals occurred with BACTEC 9240 (15 cultures) than with BACTEC 9050 (4 cultures). Likewise, more false-negative cultures occurred with BACTEC 9240 (11 cultures) than with BACTEC 9050 (8 cultures). Time to detection of positive cultures of clinically significant organisms was essentially the same with both systems with the exception of anaerobes (
N
= 10), which were recovered earlier (
P
< 0.01) with BACTEC 9240 (35.0 h) than with BACTEC 9050 (61.4 h).
Detection of Antigen in Sera of Patients with Invasive Aspergillosis: Intra- and Interlaboratory Reproducibility
Authors: Verweij Paul E. , Erjavec Zoran , Sluiters Wim , Goessens Wil , Rozenberg-Arska Marja , Debets-Ossenkopp Yvette J. , Guiot Henri F. L. , Meis Jacques F. G. M. 
Published: December 2019

ABSTRACT

The intra- and interlaboratory reproducibilities of a commercial sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of
Aspergillus
galactomannan in serum (Platelia Aspergillus; Sanofi Diagnostics Pasteur, Marnes-La-Coquette, France) were evaluated in six laboratories of university hospitals. Twenty serum samples were obtained from 12 neutropenic patients including 6 with invasive aspergillosis. These samples were blinded and sent to each center together with eight blinded ELISA-negative serum samples spiked with known concentrations of galactomannan. The centers were provided with ELISA microtiter plates from a single batch and a detailed protocol. Ten clinical samples showed ELISA reactivity, while 10 samples were ELISA negative. The mean coefficient of variation (CV) of the optical density values was 4.24% within a single assay and 25.6% between runs. The interassay CV of the ratios for the serum samples tested was 18.6%. Analysis of ordinal interpretation of the ELISA result (i.e., negative, gray zone, or positive) showed excellent reproducibility. Recalculation of the cutoff values for positive and negative samples suggested that the cutoff level recommended by the manufacturer could be lowered from 1.0 to 0.8 for negative samples and from 1.5 to 1.0 for positive samples. The intra- and interlaboratory reproducibilities were excellent when the ELISA results were interpreted as ordinal data, but considerable variation in optical density values and, to a lesser extent, in the ratios for the serum samples tested, was observed between runs. High assay variability was also found for serum samples spiked with known concentrations of galactomannan. Therefore, antigen titers in serum samples from a single patient, measured in different runs, should be compared with caution.
Microscopic Examination and Broth Culture of Cerebrospinal Fluid in Diagnosis of Meningitis
Authors: Dunbar Sherry A. , Eason Rachel A. , Musher Daniel M. , Clarridge Jill E. 
Published: December 2019

ABSTRACT

We reviewed the results of microscopic Gram stain examination and routine culture for 2,635 cerebrospinal fluid (CSF) samples processed in an adult hospital microbiology laboratory during 55 months. There were 56 instances of bacterial or fungal meningitis (16 associated with central nervous system [CNS] shunt infection), four infections adjacent to the subarachnoid space, four cases of sepsis without meningitis, and an additional 220 CSF specimens with positive cultures in which the organism isolated was judged to be a contaminant. Because 121 of these contaminants were isolated in broth only, elimination of the broth culture would decrease unnecessary work. However, 25% of the meningitis associated with CNS shunts would have been missed by this practice. The most common cause of meningitis was
Cryptococcus neoformans
, followed by
Streptococcus pneumoniae
and
Neisseria meningitidis
. In 48 of 56 (88%) of cases, examination of the Gram-stained specimen revealed the causative organism. If patients who had received effective antimicrobial therapy prior to lumbar puncture are excluded, the CSF Gram stain is 92% sensitive. Microscopic examination incorrectly suggested the presence of organisms in only 3 of 2,635 (0.1%) CSF examinations. Thus, microscopic examination of Gram-stained, concentrated CSF is highly sensitive and specific in early diagnosis of bacterial or fungal meningitis.
Use of PCR for Diagnosis of Post-Kala-Azar Dermal Leishmaniasis
Authors: Osman Omran F. , Oskam Linda , Kroon Nel C. M. , Schoone Gerard J. , Khalil El-Tahir A. G. , El-Hassan Ahmed M. , Zijlstra Ed E. , Kager Piet A. 
Published: December 2019

ABSTRACT

Microscopy and PCR were compared for use in the diagnosis of post-kala-azar dermal leishmaniasis (PKDL) in 63 patients. Aspirates of lymph nodes (samples from 52 patients), skin (23 samples), and bone marrow (18 samples) were used. For 11 patients lymph node aspiration could be repeated 6 months after they recovered from PKDL. During active PKDL, PCR was positive for 42 of 52 (80.8%) lymph node aspirates and 19 of 23 (82.7%) skin aspirates, whereas microscopy was positive for only 9 of 52 (17.3%) lymph node aspirates and 7 of 23 (30.4%) skin aspirates. PCR was always positive when parasites were seen by microscopy. When the results obtained with lymph node and skin aspirates from the same patient (
n
= 16) were compared, there was complete agreement. Bone marrow samples were negative by microscopy and PCR for 16 patients and positive by both methods for 1 patient; for one sample only the PCR was positive. PCR confirmed the co-occurrence of visceral leishmaniasis and PKDL in one patient and confirmed the suspicion of this co-occurrence in the other patient. After recovery, no parasites were found by microscopy, but 2 of 11 (18.2%) samples were still positive by PCR. Thirty negative controls were all found to be PCR negative, and 15 positive controls were all PCR positive. Cross-reactions with
Mycobacterium leprae
could be ruled out. In conclusion, PCR with inguinal lymph node or skin aspirates is suitable for confirming the clinical diagnosis of PKDL. In some patients, lymph node aspirates are probably preferred because aspiration of material from the skin may leave scars.
Cloning and Expression of the 44-Kilodalton Major Outer Membrane Protein Gene of the Human Granulocytic Ehrlichiosis Agent and Application of the Recombinant Protein to Serodiagnosis
Authors: Zhi N. , Ohashi N. , Rikihisa Y. , Horowitz H. W. , Wormser G. P. , Hechemy K. 
Published: December 2019

ABSTRACT

A 44-kDa major outer membrane protein of the human granulocytic ehrlichiosis (HGE) agent is an immunodominant antigen in human infection. A gene encoding this protein was cloned and sequenced. Southern blot results revealed the existence of multigenes homologous to the P44 gene in the genome of the HGE agent. The recombinant 44-kDa protein (rP44) was expressed by using expression vector pET30a. The reactivity of the affinity-purified rP44 was evaluated by Western immunoblot analysis and dot blot immunoassay. Western immunoblot analysis showed that mouse anti-rP44 serum reacted with 44- to 42-kDa proteins in six different HGE agent strains tested except strain 2, in which three proteins of 42, 40, and 38 kDa were recognized. Eleven HGE patient serum samples, a horse anti-HGE serum, and a horse anti-
Ehrlichia equi
serum recognized the rP44 protein. This suggests that rP44 is an HGE-
E. equi
group-specific antigen. Neither human anti-
Ehrlichia chaffeensis
serum nor rabbit anti-
Borrelia burgdorferi
serum reacted with rP44. Sera from two patients coinfected with the HGE agent and
B. burgdorferi
reacted positively with rP44 and the HGE agent. Sera from 20 HGE patients with indirect fluorescent-antibody (IFA) titers ranging from 1:20 to 1:2,560 gave distinct positive reactions in a dot immunoblot assay. There was a positive correlation between the color densities of the dot reactions and the IFA titers when greater than 50 ng of recombinant antigen per dot was used. The use of the affinity-purified rP44 protein as antigen would provide a more specific, consistent, and simpler serodiagnosis for HGE than the use of whole infected cells or purified HGE agents.
CDC Group O-3: Phenotypic Characteristics, Fatty Acid Composition, Isoprenoid Quinone Content, and In Vitro Antimicrobic Susceptibilities of an Unusual Gram-Negative Bacterium Isolated from Clinical Specimens
Authors: Daneshvar M. I. , Hill B. , Hollis D. G. , Moss C. W. , Jordan J. G. , Macgregor J. P. , Tenover F. , Weyant R. S. 
Published: December 2019

ABSTRACT

Between 1983 and 1994, 13 phenotypically similar unidentified clinical isolates were received by the Special Bacteriology Reference Laboratory, Centers for Disease Control and Prevention (CDC). Sources included blood (four strains), lung (three strains), knee fluid and duodenal tissue (one strain each), bone, and lymph node tissue (two strains each). All were aerobic glucose-oxidizing, slender, long, curved gram-negative rods that utilized xylose, sucrose, and maltose; did not grow on MacConkey agar in 1 to 2 days; were oxidase positive; hydrolyzed esculin; and grew on
Campylobacter
selective medium. All were negative for urease, indole, nitrate reduction, and gelatin hydrolysis. All were motile by means of a single polar flagellum with a noticeably short wavelength; however, motility was sometimes difficult to demonstrate. The cellular fatty acid compositions of these strains, as analyzed by gas-liquid chromatography, were unique, characterized by relatively large amounts of 16:1ω7c, 16:0, and 18:1ω7c with smaller amounts of 12:0, 3-OH-12:1, 14:0, 15:0, 18:0, Br-19:1, and 19:0cyc
11–12
. High-performance liquid chromatography and mass spectrometry of the quinone extracts of three representative strains showed ubiquinone-10 as the major component. Based on the breakpoints for the family
Enterobacteriaceae
, all the strains were susceptible in vitro to aminoglycosides, sulfamethoxazole-trimethoprim, and chloramphenicol but were resistant to most beta-lactams except imipenem. The MICs of amoxicillin-clavulanate and ciprofloxacin for these strains clustered around the breakpoints, which makes it difficult to predict the strains’ response in vivo to these agents. This group has been designated CDC oxidizer group 3 (O-3).
Quantitation of Bacteria in Blood of Typhoid Fever Patients and Relationship between Counts and Clinical Features, Transmissibility, and Antibiotic Resistance
Authors: Wain John , Diep To Song , Ho Vo Anh , Walsh Amanda M. , Hoa Nguyen Thi Tuyet , Parry Christopher M. , White Nicholas J. 
Published: December 2019

ABSTRACT

Salmonella typhi
was isolated from 369 and
Salmonella paratyphi
A was isolated from 6 of 515 Vietnamese patients with suspected enteric fever. Compared with conventional broth culture of blood, direct plating of the buffy coat had a diagnostic sensitivity of 99.5% (95% confidence interval [CI], 97.1 to 100%). Blood bacterial counts were estimated by the pour plate method. The median
S. typhi
count in blood was 1 CFU/ml (range, <0.3 to 387 CFU/ml), of which a mean of 63% (95% CI, 58 to 67%) were intracellular. The mean number of bacteria per infected leukocyte was 1.3 (interquartile range [IQR], 0.7 to 2.4) CFU/cell (
n
= 81). Children (<15 years old;
n
= 115) had higher median blood bacterial counts than adults (
n
= 262): 1.5 (range, <0.3 to 387) versus 0.6 (range, <0.3 to 17.7) CFU/ml (
P
= 0.008), and patients who excreted
S. typhi
in feces had higher bacteremias than those who did not: a median of 3 (range, <0.3 to 32) versus 1 (range, <0.3 to 68) CFU/ml (
P
= 0.02). Blood bacterial counts declined with increasing duration of illness (
P
= 0.002) and were higher in infections caused by multidrug-resistant
S. typhi
(1.3 [range, <0.3 to 387] CFU/ml;
n
= 313) than in infections caused by antibiotic-sensitive
S. typhi
(0.5 [range, <0.3 to 32] CFU/ml;
n
= 62) (
P
= 0.006). In a multivariate analysis this proved to be an independent association, suggesting a relationship between antibiotic resistance and virulence in
S. typhi.
Detection of Circulating
Paracoccidioides brasiliensis
Antigen in Urine of Paracoccidioidomycosis Patients before and during Treatment
Authors: Salina Margarete Aparecida , Shikanai-Yasuda Maria Aparecida , Mendes Rinaldo Poncio , Barraviera Benedito , Mendes Giannini Maria José Soares 
Published: December 2019

ABSTRACT

For the diagnosis and follow-up of paracoccidioidomycosis patients undergoing therapy, we evaluated two methods (immunoblotting and competition enzyme immunoassay) for the detection of circulating antigen in urine samples. A complex pattern of reactivity was observed in the immunoblot test. Bands of 70 and 43 kDa were detected more often in urine samples from patients before treatment. The immunoblot method detected gp43 and gp70 separately or concurrently in 11 (91.7%) of 12 patients, whereas the competition enzyme immunoassay detected antigenuria in 9 (75%) of 12 patients. Both tests appeared to be highly specific (100%), considering that neither fraction detectable by immunoblotting was present in urine samples from the control group. gp43 remained present in the urine samples collected during the treatment period, with a significant decrease in reactivity in samples collected during clinical recovery and increased reactivity in samples collected during relapses. Reactivity of some bands was also detected in urine specimens from patients with “apparent cure.” The detection of
Paracoccidioides brasiliensis
antigens in urine appears to be a promising method for diagnosing infection, for evaluating the efficacy of treatment, and for detecting relapse.
Invasive Infection with
Fusarium chlamydosporum
in a Patient with Aplastic Anemia
Authors: Segal Brahm H. , Walsh Thomas J. , Liu Johnson M. , Wilson Jon D. , Kwon-Chung Kyung J. 
Published: December 2019

ABSTRACT

We report the first case of invasive disease caused by
Fusarium chlamydosporum
. The patient had aplastic anemia with prolonged neutropenia and was treated with immunosuppressive therapy. While she was receiving empirical amphotericin B, a dark crusted lesion developed on her nasal turbinate. Histologic analysis revealed invasive hyaline hyphae and some darkly pigmented structures that resembled conidia of dematiaceous molds. Only after the mold was grown in culture were characteristic colonial morphology, phialides, conidia, and chlamydospores evident, thus permitting the identification of
F. chlamydosporum
. This case illustrates the ever-increasing spectrum of pathogenic
Fusarium
spp. in immunocompromised patients and emphasizes the potential pitfalls in histologic diagnosis, which may have important treatment implications.
Antimicrobial Susceptibility Testing of
Bilophila wadsworthia
Isolates Submitted for Routine Laboratory Examination
Authors: Mochida Chikako , Hirakata Yoichi , Matsuda Junichi , Iori Fumiaki , Ozaki Yumi , Nakano Michiko , Hamaguchi Koichi , Izumikawa Koichi , Yamaguchi Toshiyuki , Tomono Kazunori , Maesaki Shigefumi , Yamada Yasuaki , Kohno Shigeru , Kamihira Shimeru 
Published: December 2019

ABSTRACT

MICs of antibiotics against
Bilophila wadsworthia
isolates were measured by agar and broth microdilution with pyruvic acid and by Etest. The inoculum size influenced greatly agar dilution. Despite discrepancies in MICs depending on the measurement method used, clindamycin consistently showed potent activity. Broth microdilution and Etest appear to be candidates for laboratory susceptibility testing.
Multiplex PCR for Enterotoxigenic, Attaching and Effacing, and Shiga Toxin-ProducingEscherichia coliStrains from Calves
Authors: Franck Sophia M. , Bosworth Brad T. , Moon Harley W. 
Published: December 2019

ABSTRACTA multiplex PCR was developed to identify enterotoxigenic, attaching and effacing, and Shiga toxin-producingEscherichia colistrains by amplifying genes encoding K99 and F41 fimbriae, heat-stable enterotoxin a, intimin, and Shiga toxins 1 and 2. This multiplex PCR was specific and sensitive. It will be useful for identification ofE. colistrains which cause diarrhea in calves.
Diagnosis of
Mycobacterium microti
Infections among Humans by Using Novel Genetic Markers
Authors: van Soolingen Dick , van der Zanden Adri G. M. , de Haas Petra E. W. , Noordhoek Gerda T. , Kiers Albert , Foudraine Norbert A. , Portaels Francoise , Kolk Arend H. J. , Kremer Kristin , van Embden Jan D. A. 
Published: December 2019

ABSTRACT

As a result of DNA typing of
Mycobacterium microti
isolates from animals in the United Kingdom and The Netherlands, we diagnosed four human
M. microti
infections. These are the first
M. microti
infections among humans to be reported. Three of the patients were immunocompromised and suffered from generalized forms of tuberculosis. The fourth patient was a 34-year-old immunocompetent male with a persistent cough and undefined X-ray abnormalities. Two of the
M. microti
infections were recognized by their IS
6110
restriction fragment length polymorphism (RFLP) patterns, which showed a high degree of similarity with those of
M. microti
strains isolated from a pig and a ferret in The Netherlands. The two other human
M. microti
infections were recognized by using the recently developed DNA fingerprinting method, “spoligotyping,” directly on clinical material. All
M. microti
isolates from the United Kingdom and The Netherlands were found to contain an exceptionally short genomic direct repeat region, resulting in identical two-spacer sequence reactions in spoligotyping. In contrast, the highly similar IS
6110
RFLP patterns of the vole strains from the United Kingdom differed considerably from the RFLPs of all
M. microti
strains isolated in The Netherlands, suggesting that geographic isolation led to divergent strains in the United Kingdom and on the continent.
Determination of Types of
Enterocytozoon bieneusi
Strains Isolated from Patients with Intestinal Microsporidiosis
Authors: Liguory Olivier , David Felicia , Sarfati Claudine , Derouin Francis , Molina Jean-Michel 
Published: December 2019

ABSTRACT

To determine the types of
Enterocytozoon bieneusi
strains associated with intestinal microsporidiosis, we developed a rapid and efficient approach for typing parasites obtained from stool specimens by PCR-restriction fragment length polymorphism (PCR-RFLP). Typing was based on DNA polymorphism of the ribosomal DNA internal transcribed spacer (ITS) region of
E. bieneusi
. RFLPs generated with two restriction enzymes (
Nla
III and
Fnu
4HI) in PCR-amplified ITS products were used to classify strains into different lineages. This approach was successfully used to differentiate 78 strains that had been obtained from the stools of 65 patients with intestinal microsporidiosis. Among the 78 strains, we found four genetically unrelated lineages, showing the genetic diversity of
E. bieneusi
. Type I strains of
E. bieneusi
were found in a majority of the samples, accounting for 51 (78%) of the 65 microsporidiosis cases. In contrast, type II, III, and IV strains were found in only 8 (12%), 3 (5%), and 3 (5%) cases, respectively. All strains of
E. bieneusi
we have tested so far fall into one of four different lineages, and this study shows that human intestinal microsporidiosis is most often associated with type I strains. PCR-RFLP analysis of the ITS region of
E. bieneusi
should be useful for epidemiological studies.
Identification and Molecular Characterization of Beta-Hemolytic Streptococci Isolated from Harbor Porpoises (
Phocoena phocoena
) of the North and Baltic Seas
Authors: Swenshon M. , Lämmler C. , Siebert U. 
Published: December 2019

ABSTRACT

The present study was designed to identify and comparatively investigate 35 beta-hemolytic streptococci isolated from stranded harbor porpoises or from animals caught in fishing nets of the North and Baltic seas. According to biochemical and serological data and to lectin agglutination tests with the lectin of
Arachis hypogaea
, all 35 isolates could be classified in Lancefield’s serological group L and could be identified as
Streptococcus dysgalactiae
subsp.
dysgalactiae
. All 35 group L streptococci were uniformly sensitive to most of the antibiotics tested. To further analyze the epidemiological relationship, the isolates were subjected to macrorestriction analysis of their chromosomal DNA by pulsed-field gel electrophoresis. Digestion of the chromosomal DNA with the restriction enzymes
Sma
I and
Apa
I revealed that most of the group L streptococci seemed to be apparently identical or related. These results indicate that one clone or at least related group L streptococcal clones play an important role for infections of harbor porpoises of the North and Baltic seas. This might possibly be caused by a direct transfer of the bacteria from animal to animal.
Antigenic and Molecular Characterization of Wild Type 1 Poliovirus Causing Outbreaks of Poliomyelitis in Albania and Neighboring Countries in 1996
Authors: Fiore L. , Genovese D. , Diamanti E. , Catone S. , Ridolfi B. , Ibrahimi B. , konomi R. , van der Avoort H. G. A. M. , Hovi T. , Crainic R. , Simeoni P. , Amato C. 
Published: December 2019

ABSTRACT
Mass vaccination has led poliomyelitis to become a rare disease in a large part of the world, including Western Europe. However, in the past 20 years wild polioviruses imported from countries where polio is endemic have been responsible for outbreaks in otherwise polio-free European countries. We report on the characterization of poliovirus isolates from a large outbreak of poliomyelitis that occurred in Albania in 1996 and that also spread to the neighboring countries of Yugoslavia and Greece. The epidemics involved 145 subjects, mostly young adults, and caused persisting paralysis in 87 individuals and 16 deaths. The agent responsible for the outbreak was isolated from 74 patients and was identified as wild type 1 poliovirus by both immunological and molecular methods. Sequence analysis of the genome demonstrated the involvement of a single virus strain throughout the epidemics, and genotyping analysis showed 95% homology of the strain with a wild type 1 poliovirus strain isolated in Pakistan in 1995. Neutralization assays with both human sera and monoclonal antibodies were performed to analyze the antigenic structure of the epidemic strain, suggesting its peculiar antigenic characteristics. The presented data underline the current risks of outbreaks due to imported wild poliovirus and emphasize the need to improve vaccination efforts and also the need to implement surveillance in countries free of indigenous wild poliovirus.