bluuebunny/crossref_metadata_2025_split huggingface dataset:
Geographic Discrimination of
Paracoccidioides brasiliensis
Strains by Randomly Amplified Polymorphic DNA Analysis
Authors: Calcagno Ana María , Niño-Vega Gustavo , San-Blas Felipe , San-Blas Gioconda 
Published: December 2019

ABSTRACT

Randomly amplified polymorphic DNA (RAPD) analysis of 33
Paracoccidioides brasiliensis
strains from Argentina, Brazil, Colombia, Peru, and Venezuela produced reproducible amplification products which were sufficiently polymorphic to allow differentiation of the strains. Types generated with five primers (OPG 03, OPG 05, OPG 14, OPG 16, and OPG 18) resulted in a high discriminatory index (0.956). The discriminatory index was slightly reduced (0.940) when only two primers (OPG 3 and OPG 14) were used. A dendrogram based on these results showed a high degree of similarity among the strains, and genetic differences were expressed in clusters related to geographical regions but not to pathological features of the disease. With a few exceptions, strains were sorted into five groups by geographical origin as follows: group I, Venezuelan strains; group II, Brazilian strains; group III, Peruvian strains; group IV, Colombian strains; and group V, Argentinian strains. The group containing the most disparate strains was group V (discriminatory index, 0.633); the discriminatory index for the other four groups was 0.824. The use of primer OPG 18 by itself was sufficient to discriminate species specificity, and the use of primer OPG 14 by itself was sufficient to discriminate among the geographical locations of the strains in the sample. This method may be helpful for epidemiological studies of
P. brasiliensis.
Randomly Amplified Polymorphic DNA Genotyping of Serogroup A Meningococci Yields Results Similar to Those Obtained by Multilocus Enzyme Electrophoresis and Reveals New Genotypes
Authors: Bart Aldert , Schuurman Ilse G. A. , Achtman Mark , Caugant Dominique A. , Dankert Jacob , van der Ende Arie 
Published: December 2019

ABSTRACT

Randomly amplified polymorphic DNA (RAPD) genotyping was applied to one representative strain of each of the 84 electrophoretic types (ETs) of
Neisseria meningitidis
serogroup A previously defined by multilocus enzyme electrophoresis (MEE) (J.-F. Wang et al., Infect. Immun. 60:5267–5282, 1992). Twenty-seven additional isolates comprising six ETs were also tested. MEE and RAPD genotyping yielded similar dendrograms at the subgroup level. Similar results were obtained by both methods for 18 serogroup A meningococci isolated in The Netherlands between 1989 and 1993. Ten of these isolates defined a new subgroup, designated subgroup IX. One isolate belonged to the ET-5 complex, normally associated with serogroup B strains (D. A. Caugant et al., Proc. Natl. Acad. Sci. USA 83:4927–4931, 1986). By RAPD genotyping, meningococci can be linked to previously characterized genotypes by using a computerized database, and dendrograms based on cluster analyses can easily be generated. RAPD analysis offers advantages over MEE since intermediate numbers of isolates of serogroup A meningococci can quickly be assigned to known subgroups and new subgroups can be defined.
Monoclonal Antibody F89/160.1.5 Defines a Conserved Epitope on the Ruminant Prion Protein
Authors: O’Rourke Katherine I. , Baszler Timothy V. , Miller Janice M. , Spraker Terry R. , Sadler-Riggleman Ingrid , Knowles Donald P. 
Published: December 2019

ABSTRACT
The transmissible spongiform encephalopathies are a heterogeneous group of fatal neurodegenerative disorders occurring in humans, mink, cats, and ruminant herbivores. The occurrence of novel transmissible spongiform encephalopathies in cattle in the United Kingdom and Europe and in mule deer and elk in parts of the United States has emphasized the need for reliable diagnostic tests with standardized reagents. Postmortem diagnosis is performed by histologic examination of brain sections from affected animals. The histopathological criteria for transmissible spongiform encephalopathies include gliosis, astrocytosis, neuronal degeneration, and spongiform change. These lesions vary in intensity and anatomic location depending on the host species and genetics, stage of disease, and infectious agent source. Diagnosis by histopathology alone may be ambiguous in hosts with early cases of disease and impossible if the tissue is autolyzed. Deposition of the prion protein (an abnormal isoform of a native cellular sialoglycoprotein) in the central nervous system is a reliable marker for infection, and immunohistochemical detection of this marker is a useful adjunct to histopathology. In the present paper we describe monoclonal antibody (MAb) F89/160.1.5, which reacts with prion protein in tissues from sheep, cattle, mule deer, and elk with naturally occurring transmissible spongiform encephalopathies. This MAb recognizes a conserved epitope on the prion protein in formalin-fixed, paraffin-embedded sections after hydrated autoclaving. MAb F89/160.1.5 will be useful in diagnostic and pathogenesis studies of the transmissible spongiform encephalopathies in these ruminant species.
Typing of Human
Campylobacter jejuni
Isolates in Finland by Pulsed-Field Gel Electrophoresis
Authors: Hänninen Marja-Liisa , Pajarre Sini , Klossner Marja-Liisa , Rautelin Hilpi 
Published: December 2019

ABSTRACT

A total of 69 pulsed-field gel electrophoresis (PFGE) types were identified among 176
Campylobacter jejuni
isolates from Finnish patients. In two geographic areas studied, five predominant PFGE types comprised over 40% of the isolates. One-third of the isolates had unique PFGE types. In small outbreaks, identical PFGE patterns were demonstrated, indicating a common source of infection.
Positive Result by Serology Indicates Active
Helicobacter pylori
Infection in Patients with Atrophic Gastritis
Authors: Kokkola Arto , Rautelin Hilpi , Puolakkainen Pauli , Sipponen Pentti , Färkkilä Martti , Haapiainen Reijo , Kosunen Timo U. 
Published: December 2019

ABSTRACT

Patients with atrophic corpus gastritis and elevated
Helicobacter pylori
antibody titers but
13
C-urea breath test (
13
C-UBT) and histology results negative for
H. pylori
were randomized into eradication therapy or follow-up only. Antibody levels decreased significantly in six out of seven patients in the eradication group, while in the follow-up group, the titers declined in only one out of eight patients. In patients with atrophic corpus gastritis, positive serology results may indicate an ongoing infection in spite of negative
13
C-UBT and histology results.
Blinded, Externally Controlled Multicenter Evaluation of Light Microscopy and PCR for Detection of Microsporidia in Stool Specimens
Authors: Rinder Heinz , Janitschke Klaus , Aspöck Horst , Da Silva Alexandre J. , Deplazes Peter , Fedorko Daniel P. , Franzen Caspar , Futh Ursula , Hünger Frank , Lehmacher Anselm , Meyer Christian G. , Molina Jean-Michel , Sandfort Jörg , Weber Rainer , Löscher Thomas ,   the Diagnostic Multicenter Study Group on Microsporidia
Published: December 2019

ABSTRACT
The quality parameters for the detection of microsporidia in identical sets of 50 stool samples were determined for six laboratories where technicians used light microscopy and for six laboratories where technicians used PCR. The average overall sensitivities were 67% (89% for patient samples only) for the PCR laboratories and 54% (80% for patient samples only) for the light microscopy laboratories. Specificities were 98 and 95%, respectively. Differences in results were most apparent between the individual laboratories rather than between the two major methods used.
Detection of
Echinococcus multilocularis
in the Definitive Host: Coprodiagnosis by PCR as an Alternative to Necropsy
Authors: Dinkel Anke , von Nickisch-Rosenegk Markus , Bilger Birgit , Merli Michael , Lucius Richard , Romig Thomas 
Published: December 2019

ABSTRACT

Recently, extensions of the range of
Echinococcus multilocularis
in Europe and North America and drastic increases in fox populations in Europe put an increasing proportion of the human population at risk of alveolar echinococcosis. To obtain data on the local infection pressure, studies of the prevalence of the parasite in the animals that transmit the parasite, foxes, dogs, and cats, are urgently required. Such investigations, however, have been hampered by the need for necropsy of the host animal to specifically diagnose infection with the parasite. In this study, a nested PCR and an improved method for DNA extraction were developed to allow the sensitive and specific diagnosis of
E. multilocularis
infections directly from diluted fecal samples from foxes. The target sequence for amplification is part of the
E. multilocularis
mitochondrial 12S rRNA gene. The specificity of the method was 100% when it was tested against 18 isolates (metacestodes and adult worms) of 11 cestode species, including
E. granulosus
. The sensitivity of the method was evaluated by adding egg suspensions and individual eggs to samples of diluted feces from uninfected foxes. The presence of one egg was sufficient to give a specific signal. To confirm the PCR results, an internal probe which hybridized only with
E. multilocularis
amplification products but not with the DNA of other cestodes was constructed. In order to investigate the applicability of this method for epidemiological studies, 250 wild foxes from a area in southern Germany where echinococcosis is highly endemic were examined by both necropsy and PCR of rectal contents. The sensitivity correlated with the parasites’ number and stage of maturity. It ranged from 100% (>1,000 gravid worms) to 70% (<10 nongravid worms). On the basis of positive PCR results for 165 foxes, the sensitivity of the traditional and widely used necropsy method was found to be not higher than 76%. We therefore present this PCR system as an alternative method for the routine diagnosis of
E. multilocularis
in carnivores.
Use of Monoclonal Antibodies To Identify Serotypes of Enterovirus Isolates
Authors: Rigonan Alma S. , Mann Linda , Chonmaitree Tasnee 
Published: December 2019

ABSTRACT
Nonpoliovirus enteroviruses cause a variety of diseases that are common in young children and adults. The “gold standard” for laboratory diagnosis of enteroviruses is cell culture isolation, followed by serotype identification by neutralization assay. These procedures are time-consuming and expensive. Rapid serotype identification of enteroviruses is important in differentiating nonpoliovirus enterovirus pathogens from vaccine strain polioviruses that can be shed for some time after vaccination. In the present investigation, we evaluated a rapid method for serotype identification of enteroviruses by indirect immunofluorescence assay (IFA) using commercially available monoclonal antibodies for polioviruses, coxsackieviruses type B, and six serotypes of commonly circulating echoviruses. Of 291 isolates of enteroviruses included in the study, 95 were polioviruses and 196 were nonpoliovirus enteroviruses. Two hundred thirty-four of these (38 polioviruses and 196 nonpoliovirus enteroviruses) were consecutively grown in the laboratory over a 5-year period. IFA identified the serotypes of 74% of the consecutive isolates and 71% of all enterovirus isolates by yielding a positive staining result. The levels of agreement in the identification of the enterovirus group between IFA and neutralization tests were 92% for consecutively grown isolates and 85% for all enterovirus isolates. The sensitivity of the IFA for the detection of viruses for which specific monoclonal antibodies were applied was 73% for polioviruses, 85% for coxsackieviruses type B, and 94% for echoviruses. Specificity was near 100% for polioviruses and coxsackieviruses type B and 94% for echoviruses. We conclude that IFA can be helpful as a preliminary test for serotype identification of enteroviruses. The results are most accurate when the test identifies the isolate as a poliovirus.
Rapid Detection of
Chlamydia pneumoniae
by PCR-Enzyme Immunoassay
Authors: Jantos Christian A. , Roggendorf Rüdiger , Wuppermann Frederik N. , Hegemann Johannes H. 
Published: December 2019

ABSTRACT

Chlamydia pneumoniae
is an important human respiratory pathogen. Laboratory diagnosis of infection with this organism is difficult. To facilitate the detection of
C. pneumoniae
by PCR, an enzyme immunoassay (EIA) for analysis of PCR products was developed. Biotin-labeled PCR products generated from the 16S rRNA gene of
C. pneumoniae
were hybridized to a digoxigenin-labeled probe and then immobilized to streptavidin-coated microtiter plates. Bound PCR product-probe hybrids were detected with antidigoxigenin peroxidase conjugate and a colorimetric substrate. This EIA was as sensitive as Southern blot hybridization for the detection of PCR products and 100 times more sensitive than visualization of PCR products on agarose gels. The diagnostic value of the PCR-EIA in comparison to cell culture was assessed in throat swab specimens from children with respiratory tract infections.
C. pneumoniae
was isolated from only 1 of 368 specimens tested. In contrast, 15 patient specimens were repeatedly positive for
C. pneumoniae
by PCR and Southern analysis. All of these 15 specimens were also identified by PCR-EIA. Of the 15 specimens positive by 16S rRNA-based PCR, 13 specimens could be confirmed by
omp1
-based PCR or direct fluorescent-antibody assay. Results of this study demonstrate that PCR is more sensitive than cell culture for the detection of
C. pneumoniae
. The EIA described here is a rapid, sensitive, and simple method for detection of amplified
C. pneumoniae
DNA.
Evaluation of the VITEK 2 System for Rapid Identification of Medically Relevant Gram-Negative Rods
Authors: Funke Guido , Monnet Dominique , deBernardis Chiara , von Graevenitz Alexander , Freney Jean 
Published: December 2019

ABSTRACT

The new VITEK 2 system (bioMérieux) was evaluated at two independent sites with the identification card for gram-negative bacilli (ID-GNB card). Of the 845 strains tested, which represented 70 different taxa belonging to either the family
Enterobacteriaceae
or the nonenteric bacilli, 716 (84.7%) were correctly identified at the species level. Thirty-two (3.8%) additional strains were identified to the species level after the performance of simple, rapid manual tests (oxidase, hemolysis, indole reaction, motility, and pigmentation). For 80 (9.5%) strains, these additional tests did not lead to an identification at the species level but the correct species identification was given among the organisms listed. Only 7 (0.8%) strains were misidentified, and 10 (1.2%) were not identified. Mistakes were randomly distributed over different taxa. Due to the new, more sensitive fluorescence-based technology of the VITEK 2 system, final results were available after 3 h. Since our evaluation was mainly a stress test, it is predicted that the VITEK 2 system in conjunction with the ID-GNB card would perform well under conditions of a routine clinical laboratory in identifying members of the family
Enterobacteriaceae
and selected species of nonenteric bacteria. This system is a promising, highly automated new tool for the rapid identification of gram-negative bacilli from human clinical specimens.
Identification of a GranulocyticEhrlichiaStrain Isolated from a Horse in Switzerland and Comparison with Other Rickettsiae of theEhrlichia phagocytophilaGenogroup
Authors: Pusterla Nicola , Huder Jon B. , Feige Karsten , Lutz Hans 
Published: December 2019

ABSTRACTThis case report describes a 12-year-old Arabian mare with granulocytic ehrlichiosis. Clinical signs included fever, apathy, anorexia, icterus, limb edema, and reluctance to move. Examination of buffy coat smears revealedEhrlichiaorganisms in neutrophils and eosinophils. A band of 1,428 bp was amplified from DNA of leukocytes via nested PCR and was identified as part of theEhrlichia16S rRNA gene. It differed from the gene sequences ofEhrlichia phagocytophilaandE. equiat two and three positions, respectively. Interestingly, the nucleotide sequence of the 16S rRNA was 100% identical to that of the agent of human granulocytic ehrlichiosis.
Distribution of
Staphylococcus
Species among Human Clinical Specimens and Emended Description of
Staphylococcus caprae
Authors: Kawamura Yoshiaki , Hou Xiao-Gang , Sultana Ferdousi , Hirose Kenji , Miyake Masaki , Shu Sin-Ei , Ezaki Takayuki 
Published: December 2019

ABSTRACT

By DNA-DNA hybridization on microplates, we identified 1,230 strains of staphylococci from human clinical specimens and determined the distribution of species. The 10
Staphylococcus
species isolated most often were
S. epidermidis
(31.3%),
S. aureus
(23.3%),
S. haemolyticus
(12.2%),
S. caprae
(10.7%),
S. simulans
(4.4%),
S. hominis
(4.0%),
S. capitis
(3.9%),
S. saprophyticus
(3.6%),
S. warneri
(2.2%), and
S. lugdunensis
(1.3%). From these results, we realized that
S. caprae
strains were widely distributed in human clinical specimens. The description in
Bergey’s Manual of Systematic Bacteriology
indicates that no strains of
S. caprae
produce acid from fructose and mannitol, but all our
S. caprae
strains produced acid from fructose and mannitol. Consequently, many strains of
S. caprae
isolated from human clinical specimens have been misidentified as
S. haemolyticus
or
S. hominis
by conventional biochemical tests. In this paper, we propose an emended description of
S. caprae
.
Direct Susceptibility Testing with Positive BacT/Alert Blood Cultures by Using MicroScan Overnight and Rapid Panels
Authors: Waites Ken B. , Brookings E. S. , Moser S. A. , Zimmer B. L. 
Published: December 2019

ABSTRACT
Studies were conducted on a method of direct inoculation of MicroScan dried overnight and of rapid panels with positive aerobic blood cultures obtained from the BacT/Alert to determine antimicrobial susceptibilities. Inocula were limited to specimens that appeared unimicrobic on Gram stain. Results were compared to those obtained from panels inoculated following subculture. For 133 gram-negative bacilli, there were 94.7 and 93.5% categorical agreements between direct and standard methods for all drugs tested with overnight and rapid panels, respectively. For 104 gram-positive cocci, there were 93.2 and 93.1% categorical agreements for overnight and rapid panels, respectively. The major error (false resistance) rate for gram negatives was 1.4% for overnight versus 0.7% for rapid panels. The very major error (false susceptibility) rate was 2.7% for overnight versus 8.1% for rapid panels. The total error rates were 1.6% for overnight panels and 1.5% for rapid panels. The major error rates for gram-positive direct susceptibility tests were 2.6% for overnight and 2.5% for rapid panels. The very major error rates were 8.8 and 7.2% for overnight and rapid panels, respectively. Total error rates were 3.6% for overnight and rapid gram-positive panels. These findings suggest that susceptibility results obtained from directly inoculated gram-negative overnight panels have the greatest correlation to those obtained by standard methods. When discrepant results occur with direct-susceptibility testing, they are more likely to show false susceptibility than false resistance.
Simple, Inexpensive, Reliable Method for Differentiation of
Candida dubliniensis
from
Candida albicans
Authors: Pinjon Emmanuelle , Sullivan Derek , Salkin Ira , Shanley Diarmuid , Coleman David 
Published: December 2019

ABSTRACT

Candida dubliniensis
is a recently described pathogenic species which shares many phenotypic features with
Candida albicans
, including the ability to form germ tubes and chlamydospores. These similarities have caused significant problems in the identification of
C. dubliniensis
by the average clinical mycology laboratory. To facilitate the differentiation of these species, we investigated the growth of 120 isolates of
C. dubliniensis
and 98
C. albicans
isolates at 42 and 45°C on Emmons’ modified Sabouraud glucose agar (SGA) and 10 isolates of each species in yeast-peptone-dextrose broth. None of the
C. dubliniensis
isolates grew on the agar or in the broth medium at 45°C, while 11 isolates were capable of growing on SGA at 42°C. In contrast, all of the
C. albicans
isolates but one grew at 45°C on or in either medium. These reproducible results clearly demonstrate that the incubation of isolates suspected to be
C. dubliniensis
or
C. albicans
at 45°C provides a simple, reliable, and inexpensive method for the differentiation of the two species.
Evidence of Toscana Virus Variants Circulating in Tuscany, Italy, during the Summers of 1995 to 1997
Authors: Valassina Marcello , Cuppone Anna Maria , Bianchi Silvia , Santini Laura , Cusi Maria Grazia 
Published: December 2019

ABSTRACT
The Toscana virus can cause neurological infection in adults. This study of 112 cases of acute meningitis which occurred during the summers of 1995, 1996, and 1997 demonstrated the presence of viral RNA in the cerebrospinal fluid of 56 patients. Their sequence analysis shows four variants of the Toscana virus.
Replication of Bovine Herpesvirus Type 4 in Human Cells In Vitro
Authors: Egyed László 
Published: December 2019

ABSTRACTA reference strain (Movár 33/63) of bovine herpesvirus type 4 (BHV-4) was inoculated into 14 different human cell lines and five primary cell cultures representing various human tissues. BHV-4 replicated in two embryonic lung cell lines, MRC-5 and Wistar-38, and in a giant-cell glioblastoma cell culture. Cytopathic effect and intranuclear inclusion bodies were observed in these cells. PCR detected a 10,000-times-higher level of BHV-4 DNA. Titration of the supernatant indicated a 100-fold increase of infectious particles. Since this is the first bovine (human herpesvirus 8 and Epstein-Barr virus related) herpesvirus which replicates on human cells in vitro, the danger of possible human BHV-4 infection should not be ignored.
Diagnosis of Kala-Azar by Nested PCR Based on Amplification of the
Leishmania
Mini-Exon Gene
Authors: Katakura Ken , Kawazu Shin-Ichiro , Naya Toshimitsu , Nagakura Koichi , Ito Mamoru , Aikawa Masamichi , Qu Jing-Qi , Guan Li-Ren , Zuo Xin-Pin , Chai Jun-Jie , Chang K.-P. , Matsumoto Yoshitsugu 
Published: December 2019

ABSTRACT

To diagnose visceral leishmaniasis (kala-azar), we have developed a nested PCR method based on amplification of the mini-exon gene, which is unique and tandomly repeated in the
Leishmania
genome. Nested PCR was sufficiently sensitive for the detection of DNA in an amount equivalent to a single
Leishmania
parasite or less. We examined the usefulness of this PCR method using bone marrow aspirates and buffy coat cells collected from kala-azar patients who had or had not received chemotherapy in northwest China. We obtained PCR positivity for all of the parasitologically positive bone marrow samples from the patients. Some ambiguities with the primary PCR results were eliminated by the subsequent nested PCR. The buffy coat samples from 7 of 12 patients with splenomegaly were positive by the nested PCR, although only 2 of them were positive for parasites by culture. However, buffy coat samples from nine children, whose splenomegaly has been reduced and clinically cured by antimony treatment, were all negative. Thus, this nested PCR method represents a new tool for the diagnosis of kala-azar with patient blood samples instead of bone marrow or spleen aspirates obtained by more invasive procedures.