bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of Recombinant Diagnostic Reagents Based on pp85(U14) and p86(U11) Proteins To Detect the Human Immune Response to Human Herpesvirus 7 Infection
Authors: Stefan Alessandra , De Lillo Margherita , Frascaroli Giada , Secchiero Paola , Neipel Frank , Campadelli-Fiume Gabriella 
Published: December 2019

ABSTRACT
Human antibodies raised in response to human herpesvirus 7 (HHV-7) infection are directed predominantly to one or more HHV-7-infected cell proteins with apparent molecular masses of about 85 to 89 kDa. The genes that encode these proteins are unknown. However, several HHH-7 genes that possibly encode proteins in this molecular mass range have been identified. Thus, the proteins encoded by open reading frame U14 (85 kDa) and U11 (86 kDa) were expressed as recombinant proteins in bacteria. Of 13 human serum specimens that recognized the 85- to 89-kDa protein(s) of HHV-7-infected cells by immunoblotting, 12 were also reactive with recombinant pp85(U14) and 8 were reactive with p86(U11). It is concluded that (i) the HHV-7 immunodominant protein is pp85(U14) and (ii) the lack of posttranslational modifications in procaryotically expressed pp85 does not adversely affect the reactivity of human sera. Monoclonal antibody (MAb) 5E1 is an HHV-7-specific MAb directed to pp85(U14). Here, the HHV-7-specific epitope in pp85(U14) was finely mapped to the C′ terminal region between amino acid residues 484 and 502. However, as indicated by the low level of reactivity of human sera with the HHV-7-specific epitope recognized by MAb 5E1, human sera recognize additional epitopes of pp85(U14) that are required for their full reactivity.
rRNA Gene Internal Transcribed Spacer 1 and 2 Sequences of Asexual, Anthropophilic Dermatophytes Related to
Trichophyton rubrum
Authors: Summerbell R. C. , Haugland R. A. , Li A. , Gupta A. K. 
Published: December 2019

ABSTRACT

The ribosomal region spanning the two internal transcribed spacer (ITS) regions and the 5.8S ribosomal DNA region was sequenced for asexual, anthropophilic dermatophyte species with morphological similarity to
Trichophyton rubrum
, as well as for members of the three previously delineated, related major clades in the
T. mentagrophytes
complex. Representative isolates of
T. raubitschekii
,
T. fischeri
, and
T. kanei
were found to have ITS sequences identical to that of
T. rubrum
. The ITS sequences of
T. soudanense
and
T. megninii
differed from that of
T. rubrum
by only a small number of base pairs. Their continued status as species, however, appears to meet criteria outlined in the population genetics-based cohesion species concept of A. R. Templeton. The ITS sequence of
T. tonsurans
differed from that of the biologically distinct
T. equinum
by only 1 bp, while the ITS sequence of the recently described species
T. krajdenii
had a sequence identical to that of
T. mentagrophytes
isolates related to the teleomorph
Arthroderma vanbreuseghemii
.
Bacteremia Due to Extended-Spectrum β-Lactamase-Producing
Escherichia coli
and
Klebsiella pneumoniae
in a Pediatric Oncology Ward: Clinical Features and Identification of Different Plasmids Carrying both SHV-5 and TEM-1 Genes
Authors: Siu L. K. , Lu Po-Liang , Hsueh Po-Ren , Lin F. M. , Chang Shan-Chwen , Luh Kwen-Tay , Ho Monto , Lee Chin-Yun 
Published: December 2019

ABSTRACT

Thirteen patients who had 16 episodes of bacteremia were observed between 1993 and 1997 in a pediatric oncology ward with a high background isolation rate of cefotaxime- or aztreonam-resistant gram-negative bacteria. Four blood isolates were
Escherichia coli
and 12 were
Klebsiella pneumoniae
, and these isolates harbored extended-spectrum β-lactamases (ESBLs). All episodes of bacteremia were nosocomial, all except one of the episodes occurred in neutropenic patients, and all patients were treated with piperacillin or ceftazidime with amikacin and cefazolin prior to the onset of bacteremia. Nine of 13 patients were receiving extended-spectrum β-lactam treatment when the bacteremias caused by ESBL producers occurred. Molecular studies revealed that four
K. pneumoniae
SHV-2-producing isolates from 1994 were of the same clone. Other ESBL producers, including six that carried both TEM-1 and SHV-5, five that carried SHV-5, and one that carried SHV-2 alone, were unrelated. In conclusion, SHV-5 was present in 11 of the 16 isolates and coexisted with TEM-1 in 6 isolates. Acquisition of resistance genes probably occurred under antibiotic selection pressure. This study highlights the importance of routine checks for and detection of ESBL producers. Effective therapy against ESBL producers should be considered early for children who have malignancies and neutropenia and who are septic, despite treatment with a regimen that includes an extended-spectrum β-lactam, in a clinical setting of an increased incidence of ESBL-producing bacteria.
Detection by PCR ofNeospora caninumin Fetal Tissues from Spontaneous Bovine Abortions
Authors: Baszler Timothy V. , Gay Lawrence J. C. , Long Maureen T. , Mathison Bruce A. 
Published: December 2019

ABSTRACTThe routine diagnosis ofNeospora caninumabortion is based upon histopathologic changes in fetal tissues and identification of tissue parasites by immunohistochemistry. Confirmation ofN. caninuminfection by immunohistochemistry has low sensitivity. In the present study, we examined the utility of PCR in detectingN. caninuminfection in fetal tissues from spontaneous bovine abortion. DNA was obtained from fresh and formalin-fixed tissues from 61 bovine fetuses submitted for abortion diagnosis. Histopathology and immunohistochemistry determined the true status ofN. caninuminfection in each fetus. In formalin-fixed paraffin-embedded tissues, PCR detectedN. caninumDNA in 13 of 13 true-positive fetuses (100%) and in 1 of 16 true-negative fetuses (6%). In fresh or frozen tissues, PCR detectedN. caninumDNA in 10 of 13 true-positive fetuses (77%) and 0 of 11 true-negative fetuses (0%). PCR also detectedN. caninumDNA in 6 of 8 fetuses that had typical lesions ofN. caninumbut were immunohistochemistry negative, indicating a higher sensitivity of PCR in comparison to that of immunohistochemistry.N. caninumDNA was amplified most consistently from brain tissue. PCR detection ofN. caninumDNA in formalin-fixed, paraffin-embedded tissues was superior to that in fresh tissues, presumably because of the increased accuracy of sample selection inherent in histologic specimens.
Direct and Rapid Detection by PCR of
Erysipelothrix
sp. DNAs Prepared from Bacterial Strains and Animal Tissues
Authors: Takeshi Kouichi , Makino Souichi , Ikeda Tetsuya , Takada Noriko , Nakashiro Atsushi , Nakanishi Kazunori , Oguma Keiji , Katoh Yoshinobu , Sunagawa Hiroyuki , Ohyama Tohru 
Published: December 2019

ABSTRACT

A PCR method for rapid screening of
Erysipelothrix
spp. in the slaughterhouse was carried out by using four species-specific sets of oligonucleotide primers after initial amplification with the primer set MO101-MO102, which amplifies the 16S rRNA sequences of all four
Erysipelothrix
species. The DNA sequences coding for the rRNA gene cluster, including 16S rRNA, 23S rRNA, and the noncoding region downstream of 5S rRNA, were determined in order to design primers for the species-specific PCR detection system. The homology among the 4.5-kb DNA sequences of the rRNA genes of
Erysipelothrix rhusiopathiae
serovar 2 (DNA Data Bank of Japan accession no.
AB019247
),
E. tonsillarum
serovar 7 (accession no.
AB019248
),
E. rhusiopathiae
serovar 13 (accession no.
AB019249
), and
E. rhusiopathiae
serovar 18 (accession no.
AB019250
) ranged from 96.0 to 98.4%. The PCR amplifications were specific and were able to distinguish the DNAs from each of the four
Erysipelothrix
species. The results of PCR tests performed directly with tissue specimens from diseased animals were compared with the results of cultivation tests, and the PCR tests were completed within 5 h. The test with this species-specific system based on PCR amplification with the DNA sequences coding for the rRNA gene cluster was an accurate, easy-to-read screening method for rapid diagnosis of
Erysipelothrix
sp. infection in the slaughterhouse.
Mutation Patterns of the Reverse Transcriptase and Protease Genes in Human Immunodeficiency Virus Type 1-Infected Patients Undergoing Combination Therapy: Survey of 787 Sequences
Authors: Yahi Nouara , Tamalet Catherine , Tourrès Christian , Tivoli Natacha , Ariasi Franck , Volot Françoise , Gastaut Jean-Albert , Gallais Hervé , Moreau Jacques , Fantini Jacques 
Published: December 2019

ABSTRACTThe aim of the present study was to evaluate the resistance-associated mutations in 302 human immunodeficiency virus type 1 (HIV-1)-infected patients receiving combination therapy and monitored in Marseille, France, hospitals from January 1997 to June 1998. In the reverse transcriptase (RT) gene, the most frequent mutations were found at codons 215 (53%), 41 (34%), and 67, 70, 184, and 210 (>20%). One deletion and two insertions in the β3-β4 hairpin loop of the finger subdomain (codon 69) were detected. Interesting associations and/or exclusions of specific mutations were observed. In 96% of RT genes, a mutation at codon 70 (most frequently, K70R) was associated with a wild-type genotype at position 210 (P< 10−5). Similarly, a mutation at codon 210 (most frequently, L210W) was generally associated with mutations at codons 41 (92%) and 215 (96%) but not at codon 219 (16%) or codon 70 (4%) (P< 10−5). In the protease gene, the most prevalent mutations were at codons 63 (84%), followed by codons 10, 36, 71, 77, and 93 (ca. 20%). As for RT, pairwise associations of mutations were observed. Analysis of the mutation patterns for patients with undetectable HIV-1 loads revealed a high proportion (65%) of wild-type RT genotypes but only 18% wild-type protease genotypes. For patients with high viral loads (>100,000 copies/ml), more than 50% of the RT and protease genes displayed three or more mutations. The significant correlation between the level of viremia in plasma and the number of resistance mutations in the protease (P= 0.007) and RT (P= 0.00078) genes strengthens the importance of defining the genotype of the predominant HIV-1 quasispecies before initiating antiretroviral therapy.
Production of Monoclonal Antibodies Directed against the Microsporidium
Enterocytozoon bieneusi
Authors: Accoceberry Isabelle , Thellier Marc , Desportes-Livage Isabelle , Achbarou Abderrahim , Biligui Sylvestre , Danis Martin , Datry Annick 
Published: December 2019

ABSTRACT

Several hybridomas producing antibodies detected by indirect immunofluorescence antibody test (IFAT) were established by fusion of mouse myeloma SP2/O with spleen cells from BALB/c mice immunized against whole spores (protocol 1) or chitinase-treated spores (protocol 2) of
Enterocytozoon bieneusi
and were cloned twice by limiting dilutions. Two monoclonal antibodies (MAbs), 3B82H2 from protocol 1, isotyped as immunoglobulin M (IgM), and 6E52D9 from protocol 2, isotyped as IgG, were expanded in both ascites and culture. IFAT with the MAbs showed that both MAbs reacted exclusively with the walls of the spores of
E. bieneusi
, strongly staining the surface of mature spores, and produced titers of greater than 4,096. Immunogold electron microscopy confirmed the specific reactivities of both antibodies. No cross-reaction, either with the spores of the other intestinal microsporidium species
Encephalitozoon intestinalis
or with yeast cells, bacteria, or any other intestinal parasites, was observed. The MAbs were used to identify
E. bieneusi
spores in fecal specimens from patients suspected of having intestinal microsporidiosis. The IFAT was validated against standard staining methods (Chromotrope 2R and Uvitex 2B) and PCR. We report here the first description and characterization of two MAbs specific for the spore wall of
E. bieneusi
. These MAbs have great potential for the demonstration and species determination of
E. bieneusi
, and their application in immunofluorescence identification of
E. bieneusi
in stool samples could offer a new diagnostic tool for clinical laboratories.
Recovery of
Salmonella
by Using Selenite Brilliant Green Sulfa Enrichment Broth
Authors: Chang Chiao-tang , Yuo Chung-Yee , Shen Hui-Ching , Li A-Mai , Chen Chao-yu , Chou Jui-ling , Huang Shiao-ping 
Published: December 2019

ABSTRACT

The efficacy and sensitivity of selenite brilliant green sulfa enrichment (SBG) broth for the isolation of
Salmonella
from fecal specimens were evaluated by using both clinical and artificially infected (artificial) fecal specimens. An examination of 1,588 clinical fecal specimens found
Salmonella
in 296 specimens, including 89 cases detected by the direct-plating xylose-lysine-desoxycholate method and an additional 207 cases detected after enrichment with SBG broth. Therefore, the recovery of
Salmonella
with SBG broth is increased 3.3-fold over that by the direct-plating method alone. Furthermore, the isolation rate of
Salmonella
is higher when using SBG broth than when using gram-negative (GN) broth or GN broth supplemented with sodium selenite. To determine the sensitivity for the recovery of
Salmonella
, artificial specimens containing various amounts of
Salmonella
were prepared and analyzed. The results indicated that the sensitivity is also higher with SBG broth than with GN broth. Moreover, the optimal incubation period for SBG broth can be extended to 24 h. In conclusion, the SBG enrichment method provides a higher recovery rate of
Salmonella
from fecal specimens.
Improved Diagnosis of
Trichomonas vaginalis
Infection by PCR Using Vaginal Swabs and Urine Specimens Compared to Diagnosis by Wet Mount Microscopy, Culture, and Fluorescent Staining
Authors: van der Schee Cindy , van Belkum Alex , Zwijgers Lisette , van der Brugge Esther , O'neill Errol L. , Luijendijk Ad , van Rijsoort-Vos Tineke , van der Meijden Willem I. , Verbrugh Henri , Sluiters Hans J. F. 
Published: December 2019

ABSTRACT

Four vaginal cotton swab specimens were obtained from each of 804 women visiting the outpatient sexually transmitted disease clinic of the Erasmus University Medical Center Rotterdam, Rotterdam, The Netherlands, for validation of various forms of
Trichomonas vaginalis
diagnostic procedures. One swab specimen was immediately examined by wet mount microscopy, a second swab was placed in Kupferberg's Trichosel medium for cultivation, and two swabs were placed in phosphate-buffered saline (PBS), pH 7.2. The resulting PBS suspension was used for direct staining with acridine orange and fluorescence microscopy, inoculation of modified Diamond's culture medium, and a PCR specific for
T. vaginalis
. A total of 70 samples positive in one or more of the tests were identified: 31 (3.8%) infections were detected by wet mount microscopy, and 36 (4.4%) were identified by acridine orange staining, as opposed to 40 (4.9%) and 46 (5.7%) positives in modified Diamond's and Trichosel media, respectively. PCR was positive for 61 (7.5%) samples. Secondly, from each of 200 women were obtained a urine sample and a vaginal cotton swab specimen, and 200 urine samples were obtained from men. For the women, 15 (7.4%) of the samples showed a positive result for either the wet mount (
n
= 1), Trichosel culture (
n
= 6), PCR on the vaginal swab sample (
n
= 10), or PCR on the urine specimen (
n
= 11). Four men (2%) were diagnosed with a
T. vaginalis
infection. Thus, PCR appears to be the method of choice for the detection of genital infections with
T. vaginalis.
Mycotic Keratitis Due toCurvularia senegalensisand In Vitro Antifungal Susceptibilities ofCurvulariaspp
Authors: Guarro Josep , Akiti Tiyomi , Horta Roberto Almada- , Morizot Leite-Filho L. A. , Gené Josepa , Ferreira-Gomes Sueli , Aguilar Carme , Ortoneda Montserrat 
Published: December 2019

ABSTRACTA case of mycotic keratitis due toCurvularia senegalensisis reported. This case represents the third known reported infection caused by this rare species. Fungal hyphae were detected in corneal scrapings, and repeated cultures were positive for this fungi. The patient was presumed cured after a corneal transplant and treatment with itraconazole, but the infection recurred and the patient is waiting for a keratoplasty. The in vitro antifungal susceptibilities of the case strain and another 24 strains belonging to seven species ofCurvulariawere tested for six antifungal agents. With the exception of flucytosine, and occasionally fluconazole, the other drugs assayed (amphotericin B, miconazole, itraconazole, and ketoconazole) were highly effective in vitro.
Lumbar Vertebral Osteomyelitis with Mycotic Abdominal Aortic Aneurysm Caused by Highly Penicillin-Resistant
Streptococcus pneumoniae
Authors: Naktin Jaan , DeSimone Joseph 
Published: December 2019

ABSTRACT

We present a case of vertebral osteomyelitis with an adjacent abdominal aortic mycotic aneurysm caused by a highly penicillin-resistant
Streptococcus pneumoniae
strain. The occurrence of all three phenomena in a single patient has not been previously described. This presentation offers the opportunity to reflect on the increasing incidence of
S. pneumoniae
as a resistant pathogen, the treatment of highly penicillin-resistant
S. pneumoniae
, and the etiologic agents of both vertebral osteomyelitis and mycotic aneurysm.
New PCR Primer Pairs Specific for
Cryptococcus neoformans
Serotype A or B Prepared on the Basis of Random Amplified Polymorphic DNA Fingerprint Pattern Analyses
Authors: Aoki Francisco Hideo , Imai Tamae , Tanaka Reiko , Mikami Yuzuru , Taguchi Hideaki , Nishimura Nancy Fusae , Nishimura Kazuko , Miyaji Makoto , Schreiber Angelica Zaninella , Branchini Maria Luiza Moretti 
Published: December 2019

ABSTRACT

Thirty-three strains of
Cryptococcus neoformans
were isolated from clinical specimens, including specimens from AIDS patients in Brazil, and were classified into two serotypes; we detected 31 and 2 strains of serotypes A and B, respectively. Random amplified polymorphic DNA (RAPD) fingerprint pattern analyses of these strains of serotypes A and B showed that the patterns were similar for strains of each serotype when three 10-mer primers were used as the RAPD primers. Comparative studies of the fingerprint patterns of the study isolates with those of the reference strains also showed that the RAPD patterns for strains of each serotype were related and that most of the fingerprint bands existed commonly for all strains of each serotype tested. The common RAPD bands (an approximately 700-bp band for serotype A and an approximately 450-bp band for serotype B) were extracted and the DNA sequences were determined. Using this information, we prepared two and one PCR primer pairs which were expected to be specific for
C. neoformans
serotypes A and B, respectively. Use of each PCR primer combination thus prepared for serotype A or B was 100% successful in identifying the respective
C. neoformans
serotypes, including the 33 clinical isolates tested in the present study. Among these combinations, one for serotype A was found to amplify DNA from
C. neoformans
serotype B as well as serotype A. Serotype B-specific PCR primer pairs amplified DNA from not only serotype B strains but also from serotype C strains. The usefulness of other serotype-specific PCR primers for clinical
C. neoformans
isolates is discussed.
Flow Cytometric Analysis of Microsporidia Belonging to the Genus
Encephalitozoon
Authors: Moss Delynn M. , Croppo Gian P. , Wallace Sara , Visvesvara Govinda S. 
Published: December 2019

ABSTRACT

Flow cytometry was used in the identification of human microsporidia belonging to the genus
Encephalitozoon
. Microsporidian spores of
Encephalitozoon hellem
,
E. cuniculi
, and
E. intestinalis
were propagated in axenic cultures of monkey kidney E6 cells, purified with Percoll, and exposed to homologous and heterologous rabbit antiserum and monoclonal antibody prepared against
E. hellem
spores. After reaction to goat anti-rabbit immunoglobulin G (IgG) or goat anti-mouse IgG conjugated to fluorescein isothiocyanate, fluorescence histograms from gated data on light-scatter profiles showed that rabbit anti-
E. hellem
serum was reactive to
E. hellem
spores but also had cross-reactivity to spores of
E. cuniculi
and
E. intestinalis
. On the other hand, fluorescence histograms showed that rabbit anti-
E. cuniculi
and rabbit anti-
E. intestinalis
sera were reactive with homologous spores only. Monoclonal antibody prepared against
E. hellem
reacted only with spores of
E. hellem
. Neither the polyclonal antibodies nor the monoclonal antibodies reacted with
Cryptosporidium parvum
oocysts. Fluorescence histograms of spores treated with 10% formalin also showed reactivity, but the number of events in the most intense peaks of fluorescence was fewer (7 to 42%, depending on species) than the number of events in the most intense peaks of fluorescence for nontreated spores. By flow cytometry, formalin-treated and nontreated spores of
Encephalitozoon
were identified to the species level by using gated data on light-scatter profiles and analyzing the fluorescence histograms from the indirect immunofluorescence of the spores. Once a procedure is established for the isolation of
Encephalitozoon
spores from clinical specimens, identification of spores by flow cytometry may be useful not only for diagnosis but also for epidemiologic studies.
Detection of Rubella Virus-Specific Immunoglobulin G in Saliva by an Amplification-Based Enzyme-Linked Immunosorbent Assay Using Monoclonal Antibody to Fluorescein Isothiocyanate
Authors: Vyse A. J. , Brown D. W. G. , Cohen B. J. , Samuel R. , Nokes D. J. 
Published: December 2019

ABSTRACT
An immunoglobulin G (IgG)–capture enzyme-linked immunosorbent assay (ELISA) for rubella virus is described. The assay uses a fluorescein isothiocyanate (FITC)–anti-FITC amplification system. The detection limit of the ELISA was approximately 7 IU of rubella virus-specific IgG per ml of serum sample. For saliva samples the performances of the capture ELISA and previously described radioimmunoassay were assessed, and the results of those two assays were compared to the rubella virus-specific IgG result obtained by a commercial ELISA (Behring Enzygnost) with a panel of paired serum and saliva samples. This comparison showed that the capture ELISA with saliva was more sensitive than the radioimmunoassay and that the results correlated better with the serum IgG result than the results of the radioimmunoassay did, with an overall sensitivity of 82% and a rank correlation of 0.68, whereas the sensitivity and rank correlation for the radioimmunoassay were 74% and 0.45, respectively. For subjects of 10 years of age or younger, the ELISA with saliva had a sensitivity of 94% and a specificity of 100% compared to the results of the ELISA (Behring Enzygnost) for rubella virus-specific IgG with corresponding serum samples. The sensitivity was much lower for subjects ages 17 years or older. The assay may have wider epidemiological use with saliva specimens, particularly those from children.
Quantitation of Human Immunodeficiency Virus Type 2 DNA in Peripheral Blood Mononuclear Cells by Using a Quantitative-Competitive PCR Assay
Authors: Gomes Perpétua , Taveira Nuno Costa , Pereira JoséMoniz , Antunes Francisco , Ferreira M. O. Santos , Lourenço Maria Helena 
Published: December 2019

ABSTRACT

A new quantitative-competitive PCR-based human immunodeficiency virus type 2 (HIV-2) proviral DNA assay (QC-PCR) was developed and used to determine the proviral load in HIV-2-infected individuals. Proviral load varied considerably, with means of 1,831 copies per 10
6
peripheral blood mononuclear cells for asymptomatic subjects (
n
= 19) and 2,587 for AIDS patients (
n
= 2). HIV-2 viral and proviral loads also varied significantly over time in asymptomatic patients. These data suggest that a high level of virus replication occurs throughout the asymptomatic phase of HIV-2 infection.