bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fatal Case of
Trichoderma harzianum
Infection in a Renal Transplant Recipient
Authors: Guarro Josep , Antolín-Ayala María Isabel , Gené Josepa , Gutiérrez-Calzada Jesús , Nieves-Díez Carlos , Ortoneda Montserrat 
Published: December 2019

ABSTRACT

We describe the second known case of human infection by
Trichoderma harzianum
. A disseminated fungal infection was detected in the postmortem examination of a renal transplant recipient and confirmed in culture. The only other reported infection by this fungus caused peritonitis in a diabetic patient. The in vitro antifungal susceptibilities of the clinical strain and three other strains of
Trichoderma
species to six antifungal drugs are provided. This case illustrates the widening spectrum of opportunistic
Trichoderma
spp. in immunocompromised patients and emphasizes the problems in diagnosing invasive fungal diseases.
Septicemia in Neutropenic Patients Infected with
Clostridium tertium
Resistant to Cefepime and Other Expanded-Spectrum Cephalosporins
Authors: Steyaert Sophia , Peleman Renaat , Vaneechoutte Mario , De Baere Thierry , Claeys Geert , Verschraegen Gerda 
Published: December 2019

ABSTRACT

Clostridium tertium
was isolated from two immunocompromised patients with septicemia, fever, and gastrointestinal symptoms. The strains were resistant to ceftazidime, cefepime, and clindamycin; intermediately resistant to penicillin; and susceptible to metronidazole, quinolones, and vancomycin.
Identity of a Novel Swine Hepatitis E Virus in Taiwan Forming a Monophyletic Group with Taiwan Isolates of Human Hepatitis E Virus
Authors: Hsieh Sen-Yung , Meng Xiang-Jin , Wu Ying-Hua , Liu Shih-Tung , Tam Albert W. , Lin Dneg-Yn , Liaw Yun-Fan 
Published: December 2019

ABSTRACT

Recently, we found that more than 10% of the cases of acute non-A, non-B, non-C hepatitis in Taiwan were caused by a novel strain of hepatitis E virus (HEV). Since none of these patients had a history of travel to areas where HEV is endemic, the source of transmission remains unclear. The recent discovery of a swine HEV in herd pigs in the United States has led us to speculate that HEV may also circulate in herd pigs in Taiwan and may serve as a reservoir for HEV in Taiwan. Of 275 herd pigs obtained from 10 pig farms in Taiwan, 102 (37%) were seropositive for serum anti-HEV immunoglobulin G (IgG). A 185-bp genomic sequence within the ORF-2 of the HEV genome was amplified and cloned from serum samples of an anti-HEV positive pig and subsequently from serum samples of a patient with acute hepatitis E. Sequence comparison revealed that the swine and human isolates of HEV share 97.3% identity. Phylogenetic analyses further showed that the Taiwan swine and human isolates of HEV form a distinct branch divergent from all other known strains of HEV, including the U.S. swine strain. To examine the potential risk of cross-species transmission of swine HEV to humans, the seroprevalences of anti-HEV IgG in 30 swine handlers, 20 pork dealers, and 50 control subjects were assessed and were found to be 26.7, 15, and 8%, respectively (for swine handlers versus controls,
P
= 0.048). Our findings may help provide an understanding of the modes of HEV transmission and may also raise potential public health concerns for HEV zoonosis.
Specific Detection of
Aspergillus
Species in Blood and Bronchoalveolar Lavage Samples of Immunocompromised Patients by Two-Step PCR
Authors: Skladny Heyko , Buchheidt Dieter , Baust Corinna , Krieg-Schneider Frank , Seifarth Wolfgang , Leib-Mösch Christine , Hehlmann Rüdiger 
Published: December 2019

ABSTRACT

The increasing incidence of aspergillosis, a life-threatening infection in immunocompromised patients, emphasizes the need to improve the currently limited diagnostic tools. We developed a two-step PCR assay that specifically amplifies a region of the 18S rRNA gene that is highly conserved in
Aspergillus
species. A number of primers with the least homology to equivalent human or
Candida
gene sequences were screened for the pairs that gave the highest sensitivity and specificity. No cross-reaction with the wide range of fungal and bacterial pathogens so far tested was observed. This assay allows direct and rapid detection of down to 10 fg of
Aspergillus
DNA corresponding to 1 to 5 CFU per ml of blood. A total of 315 blood and bronchoalveolar lavage samples from 140 subjects, including 93 patients at risk for invasive fungal disease, were screened. The result was a 100% correlation between positive histology, culture, or high-resolution computed tomography findings and PCR results. The test specificity was 89%. Our data point to the considerable potential clinical value of this simple, specific, rapid, and inexpensive PCR assay for improving the means of early diagnosis of systemic aspergillosis in high-risk patients.
Western and Dot Blotting Analyses ofEhrlichia chaffeensisIndirect Fluorescent-Antibody Assay-Positive and -Negative Human Sera by Using Native and RecombinantE. chaffeensisandE. canisAntigens
Authors: Unver Ahmet , Rikihisa Yasuko , Ohashi Norio , Cullman Louis C. , Buller Richard , Storch Gregory A. 
Published: December 2019

ABSTRACTHuman monocytic ehrlichiosis is an emerging infectious disease caused byEhrlichia chaffeensis, a gram-negative obligatory intracellular bacterium closely related toE. canis. The immunoreactive recombinant fusion proteins rP28 and rP30 have become available after cloning and expressing of the 28- and 30-kDa major outer membrane protein genes ofE. chaffeensisandE. canis, respectively. Western immunoblotting was performed to analyze the antibody responses of the 37E. chaffeensisindirect fluorescent-antibody assay (IFA)-positive and 20 IFA-negative serum specimens with purified whole organisms, rP28, and rP30. All IFA-negative sera were negative with purified whole organisms, rP28, or rP30 by Western immunoblot analysis (100% relative diagnostic specificity). Of 37 IFA-positive sera, 34 sera reacted with any native proteins ofE. chaffeensisranging from 44 to 110 kDa, and 30 sera reacted with 44- to 110-kDa nativeE. canisantigens. The 28-kDaE. chaffeensisand 30-kDaE. canisnative proteins were recognized by 25 IFA-positive sera. Fifteen IFA-positive sera reacted with rP28 by Western blot analysis, whereas 34 IFA-positive sera reacted with rP30 (92% relative diagnostic specificity), indicating that rP30 is more sensitive than rP28 for detecting the antibodies in IFA-positive sera. These 34 IFA-positive sera were positive by the dot blot assay with rP30, distinguishing them from IFA-negative sera. Except for three rP30-negative but IFA-positive specimens that instead showed anE. ewingiiinfection-like profile by Western immunoblotting, the results of Western and dot blot assays with rP30 matched 100% with the IFA test results. Densitometric analysis of dot blot reactions showed a positive correlation between the dot density and the IFA titer. These results suggest that rP30 antigen would provide a simple, consistent, and rapid serodiagnosis for human monocytic ehrlichiosis.
Multiplex PCR for Detection and Typing of Porcine Circoviruses
Authors: Ouardani M. , Wilson L. , Jetté R. , Montpetit C. , Dea S. 
Published: December 2019

ABSTRACT
Sets of oligonucleotide primers were designed according to the sequences of the open reading frames (ORFs) ORF1 and ORF2 of the prototype nonpathogenic PK-15 strain of porcine circovirus (PCV) type 1 (PCV-1). By the PCR performed with the various primer sets, genomic DNA or RNA from other bacterial or viral pathogens of the respiratory tracts of pigs could not be amplified. A positive amplification reaction could be visualized with DNA extracted from a viral suspension containing as few as 10 viral particles per ml. No DNA fragment could be amplified from lysates of continuous porcine cell lines (PT, ST, and PFT cells) known to be negative for PCV. When tested with clinical samples from pigs, the results of the single PCR method showed nearly 93% (13 of 14 samples) correlation with histopathological and immunohistochemical findings. Interestingly, subclinical PCV infections could be detected by single PCR with clinical samples that have been submitted from animals with irrelevant cases of respiratory and/or enteric problems. On the basis of the nucleotide sequences of PCV strains (PCV-2) recently associated with outbreaks of postweaning multisystemic wasting syndrome (PWMS) in Quebec, Canada, pig farms, other primers were designed from the PCV-1 genome, and these primers failed to amplify genomic fragments specific to the ORF1 or ORF2 genes of clinical isolates associated with PWMS but amplified DNA from the PCV-1 strain. Two rapid multiplex PCR (mPCR) methods have been developed to distinguish between both genotypes of PCV. By those two mPCR methods, (i) species-specific primer pairs were used to amplify a DNA fragment of 488 bp specific for the ORF2 genes of both genotypes, whereas a 375-bp fragment was amplified from the ORF1 gene of the PCV-1 strain only, or (ii) species-specific primer pairs were used to amplify a DNA fragment of 646 bp specific for the ORF1 genes of both genotypes, whereas a 425-bp fragment was amplified from the ORF2 gene of the PCV-1 strain only. By both mPCR methods, a PCV-2 infection was demonstrated in tissues of 94.2% (33 of 35) of the sick pigs tested, in agreement with previous findings showing the close association of this new genotype of PCV with outbreaks of PMWS in Europe and North America. On the other hand, a PCV-1 infection was confirmed in only 5.7% (2 of 35) of the pigs, and confirmation of a mixed infection with PCV-2 was obtained by a single PCR with PCV-2-specific primers.
Comparison of Four Clinical Specimen Types for Detection of Influenza A and B Viruses by Optical Immunoassay (FLU OIA Test) and Cell Culture Methods
Authors: Covalciuc Kristi A. , Webb Kenneth H. , Carlson Curtis A. 
Published: December 2019

ABSTRACT
Although laboratory diagnosis of respiratory viruses has been widely studied, there is a relative insufficiency of literature examining the impact of specimen type on the laboratory diagnosis of influenza A and B. In a clinical study comparing the FLU OIA test with 14-day cell culture, clinical specimens from nasopharyngeal swabs, throat swabs, nasal aspirates, and sputum were obtained from patients experiencing influenza-like symptoms. A total of 404 clinical specimens were collected from 184 patients. Patients were defined as influenza positive if the viral culture of a specimen from any sample site was positive. Patients were defined as influenza negative if the viral cultures of specimens from all sample sites were negative. By this gold standard, culture and FLU OIA test results for each sample type were compared. For each of the four specimen types, the viral culture and FLU OIA test demonstrated equal abilities to detect the presence of influenza A or B virus or viral antigen. Sputum and nasal aspirate samples were the most predictive of influenza virus infection. Throat swabs were the least predictive of influenza virus infection, with both tests failing to detect influenza virus in nearly 50% of the throat samples studied.
Transmission Dynamics of Tuberculosis in a High-Incidence Country: Prospective Analysis by PCR DNA Fingerprinting
Authors: Haas W. H. , Engelmann G. , Amthor B. , Shyamba S. , Mugala F. , Felten M. , Rabbow M. , Leichsenring M. , Oosthuizen O. J. , Bremer H. J. 
Published: December 2019

ABSTRACT

We have prospectively analyzed the DNA fingerprints of
Mycobacterium tuberculosis
strains from a random sample of patients with newly diagnosed tuberculosis in Windhoek, Namibia. Strains from 263 smear-positive patients in whom tuberculosis was diagnosed during 1 year were evaluated, and the results were correlated with selected epidemiological and clinical data. A total of 163 different IS
6110
fingerprint patterns were observed among the 263 isolates. Isolates from a high percentage of patients (47%) were found in 29 separate clusters, with a cluster defined as isolates with 100% matching patterns. The largest cluster included isolates from 39 patients. One predominant strain of
M. tuberculosis
caused 15% of cases of smear-positive pulmonary tuberculosis in Windhoek. That strain was also prevalent in the north of the country, suggesting that in contrast to other African countries with isolates with high levels of diversity in their DNA fingerprint patterns, only a restricted number of different strains significantly contribute to the tuberculosis problem in Namibia.
Confirmation of Low-Titer, Herpes Simplex Virus-Positive Specimen Results by the Enzyme-Linked Virus-Inducible System (ELVIS) Using PCR and Repeat Testing
Authors: Patel Navin , Kauffmann Lynn , Baniewicz Geri , Forman Michael , Evans Martin , Scholl David 
Published: December 2019

ABSTRACT

The ELVIS HSV
Id
test kit (an enzyme-linked virus-inducible system) (Diagnostic Hybrids, Inc.) uses genetically engineered BHK cells to produce a detectable enzyme, beta-galactosidase, upon infection with either herpes simplex virus (HSV) type 1 (HSV-1) or HSV-2. Twenty six ELVIS-positive clinical specimens were selected for study by PCR and with monoclonal antibodies because they were originally low-titer HSV-positive specimens by ELVIS but HSV antibody nonreactive upon follow-up staining of the ELVIS monolayer. Twenty-one of 26 specimens were frozen, thawed, and retested with ELVIS without removing the cellular debris from the specimen; 18 were ELVIS positive and 3 were ELVIS negative on retesting. A typing result was provided upon retesting for 14 of 18 ELVIS-positive specimens (11 were HSV-1 and 3 were HSV-2) with HSV-specific monoclonal antibodies; no antibody signal was observed for 4 of 18 ELVIS-positive specimens. Sixteen of 26 specimens were subjected to blinded PCR analysis with two different primer sets, including all those that were repeat tested with ELVIS without success and those that had insufficient quantity for repeat testing. All 16 specimens analyzed were PCR positive with primer set 1; 15 of 16 were also positive with primer set 2, with the HSV type identified for all specimens (7 were HSV-1 and 8 were HSV-2). These results indicate that the original ELVIS result with these low-titer specimens was correct and further confirm the sensitivity and specificity of ELVIS HSV
Id
as a rapid, cell culture-based kit for the detection of HSV.
Sensitive and Specific Serodiagnosis of Lyme Disease by Enzyme-Linked Immunosorbent Assay with a Peptide Based on an Immunodominant Conserved Region ofBorrelia burgdorferiVlsE
Authors: Liang Fang Ting , Steere Allen C. , Marques Adriana R. , Johnson Barbara J. B. , Miller James N. , Philipp Mario T. 
Published: December 2019

ABSTRACTVlsE, the variable surface antigen ofBorrelia burgdorferi, contains an immunodominant conserved region named IR6. In the present study, the diagnostic performance of a peptide enzyme-linked immunosorbent assay (ELISA) based on a 26-mer synthetic peptide (C6) with the IR6sequence was explored. Sensitivity was assessed with serum samples (n= 210) collected from patients with clinically defined Lyme disease at the acute (early localized or early disseminated disease), convalescent, or late disease phase. The sensitivities for acute-, convalescent-, and late-phase specimens were 74% (29 of 39), 85 to 90% (34 of 40 to 35 of 39), and 100% (59 of 59), respectively. Serum specimens from early neuroborreliosis patients were 95% positive (19 of 20), and those from an additional group of patients with posttreatment Lyme disease syndrome yielded a sensitivity of 62% (8 of 13). To assess the specificity of the peptide ELISA, 77 serum samples from patients with other spirochetal or chronic infections, autoimmune diseases, or neurologic diseases and 99 serum specimens from hospitalized patients in an area where Lyme disease is not endemic were examined. Only two potential false positives from the hospitalized patients were found, and the overall specificity was 99% (174 of 176). Precision, which was assessed with a panel of positive and negative serum specimens arranged in blinded duplicates, was 100%. Four serum samples with very high anti-OspA antibody titers obtained from four monkeys given the OspA vaccine did not react with the C6peptide. This simple, sensitive, specific, and precise ELISA may contribute to alleviate some of the remaining problems in Lyme disease serodiagnosis. Because of its synthetic peptide base, it will be inexpensive to manufacture. It also will be applicable to serum specimens from OspA-vaccinated subjects.
Predominant
Staphylococcus aureus
Isolated from Antibiotic-Associated Diarrhea Is Clinically Relevant and Produces Enterotoxin A and the Bicomponent Toxin LukE-LukD
Authors: Gravet Alain , Rondeau Murielle , Harf-Monteil Colette , Grunenberger Fabienne , Monteil Henri , Scheftel Jean-Michel , Prévost Gilles 
Published: December 2019

ABSTRACT

Staphylococcus aureus
was isolated as the predominant or only isolate from cultures of stools of 60 patients over 2 years in a university hospital, leading to the collection of 114 isolates. Diarrhea was observed in 90% of the patients. Ninety-eight percent of the patients had received antibiotics in the month before the diarrhea. Ninety-two percent of the
S. aureus
isolates were methicillin resistant.
S. aureus
was encountered with antibiotic-associated diarrhea among 47 quite elderly patients affected or not affected by a gastrointestinal disease. Among the antimicrobial treatments, cessation of the previous therapy when possible or rapid application of oral vancomycin therapy was the most appropriate. Analysis of total DNA by pulsed-field gel electrophoresis revealed 27 different
Sma
I pulsotypes distributed in 15 clusters. The pulsotypes never differed for related isolates from a single patient, even if they originated from patients with bacteremia.
S. aureus
was not isolated as the predominant isolate in cultures of stools of 57 patients who received an antimicrobial treatment for more than 5 days without diarrhea. Occurence of production of both enterotoxin A and the bicomponent leucotoxin LukE-LukD by the
S. aureus
isolates was significantly different from that by random isolates. The results strongly suggest that when predominant in stool samples,
S. aureus
should be considered a possible etiologic agent for some cases of antibiotic-associated diarrhea.
Performance Criteria of DNA Fingerprinting Methods for Typing ofHelicobacter pyloriIsolates: Experimental Results and Meta-Analysis
Authors: Burucoa Christophe , Lhomme Vincent , Fauchere Jean Louis 
Published: December 2019

ABSTRACTTyping systems are used to discriminate between isolates ofHelicobacter pylorifor epidemiological and clinical purposes. Discriminatory power and typeability are important performance criteria of typing systems. Discriminatory power refers to the ability to differentiate among unrelated isolates; it is quantitatively expressed by the discriminatory index (DI). Typeability refers to the ability of the method to provide an unambiguous result for each isolate analyzed; it is quantitatively expressed by the percentage of typeable isolates. We evaluated the discriminatory power and the typeability of the most currently used DNA fingerprinting methods for the typing ofH. pyloriisolates: ribotyping, PCR-based restriction fragment length polymorphism (PCR-RFLP) analysis, and random amplified polymorphism DNA (RAPD) analysis. Forty epidemiologically unrelated clinical isolates were selected to constitute a test population adapted to the evaluation of these performance criteria. A meta-analysis of typeability and discriminatory power was conducted retrospectively with raw data from published studies in which ribotyping, PCR-RFLP, RAPD, repetitive extragenic palindromic DNA sequence-based PCR (REP-PCR), or pulsed-field gel electrophoresis (PFGE) was used. Experimental results and the meta-analysis demonstrated the optimal typeability (100%) and the excellent discriminatory powers of PCR-based typing methods: RAPD analysis, DIs, 0.99 to 1; REP-PCR, DI, 0.99; and PCR-RFLP analysis, DIs, 0.70 to 0.97). Chromosome restriction-based typing methods (ribotyping and PFGE) are limited by a low typeability (12.5 to 75%) that strongly decreases their discriminatory powers: ribotyping, DI, 0.92; PFGE, DIs, 0.24 to 0.88. We do not recommend the use of ribotyping and PFGE for the typing ofH. pyloriisolates. We recommend the use of PCR-based methods.
Characteristics of
Streptococcus pyogenes
Serotype M1 and M3 Isolates from Patients in Japan from 1981 to 1997
Authors: Murase Toshiyuki , Suzuki Rieko , Osawa Ro , Yamai Shiro 
Published: December 2019

ABSTRACT

Streptococcus pyogenes
isolates obtained in 1981 to 1997 from patients and healthy subjects were characterized by pulsed-field gel electrophoresis (PFGE) patterns, biotyping, and the presence of
spe
genes encoding streptococcal pyrogenic exotoxins. Changes in the profiles were shown in the serotype M1/T1 isolates from pharyngitis over this period, but not in serotype M3/T3 isolates. The characteristics of isolates from patients with toxic shock-like syndrome (TSLS) were comparable to those of the other isolates, including those from healthy subjects. This finding suggests that further phenotypic and molecular characterization, such as investigating the genomic difference represented by the pathogenicity island, of isolates with apparently the same profiles would be necessary to determine the etiology of diseases caused by
S. pyogenes
, including TSLS.
Unusual, High Genetic Diversity of Aleutian Mink Disease Virus
Authors: Olofsson Anders , Mittelholzer Christian , Treiberg Berndtsson Louise , Lind Lars , Mejerland Torbjörn , Belák Sándor 
Published: December 2019

ABSTRACT
The genetic diversity of Aleutian mink disease virus (AMDV) was examined. Sequences obtained from 35 clinical samples were compared with five published sequences. An unusual, high genetic variability was revealed. Three phylogenetic subgroups of AMDV were identified, and the presence of more than one genotype at some farms was detected.
Rapid Identification of
Campylobacter
,
Arcobacter
, and
Helicobacter
Isolates by PCR-Restriction Fragment Length Polymorphism Analysis of the 16S rRNA Gene
Authors: Marshall Stephen M. , Melito Pasquale L. , Woodward David L. , Johnson Wendy M. , Rodgers Frank G. , Mulvey Michael R. 
Published: December 2019

ABSTRACT

A rapid two-step identification scheme based on PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the 16S rRNA gene was developed in order to differentiate isolates belonging to the
Campylobacter
,
Arcobacter
, and
Helicobacter
genera. For 158 isolates (26 reference cultures and 132 clinical isolates), specific RFLP patterns were obtained and species were successfully identified by this assay.
Molecular Epidemiology of a Cluster of Cases Due to
Klebsiella pneumoniae
Producing SHV-5 Extended-Spectrum β-Lactamase in the Premature Intensive Care Unit of a Hungarian Hospital
Authors: Szabó Dóra , Filetóth Zsolt , Szentandrássy Julianna , Némedi Mária , Tóth Erzsébet , Jeney Csaba , Kispál Gyula , Rozgonyi Ferenc 
Published: December 2019

ABSTRACT

Fifteen nosocomial cases of extended-spectrum β-lactamase-producing
Klebsiella pneumoniae
occurred among 132 neonates in a premature intensive care unit in Hungary in June through November 1998. Fourteen strains were indistinguishable by molecular biological typing and harbored the same single conjugative extended-spectrum β-lactamase-encoding plasmid that was spontaneously found in a
Serratia marcescens
strain in the same patient.