bluuebunny/crossref_metadata_2025_split huggingface dataset:
Coinfection with Multiple Tick-Borne Pathogens in a Walker Hound Kennel in North Carolina
Authors: Kordick S. K. , Breitschwerdt E. B. , Hegarty B. C. , Southwick K. L. , Colitz C. M. , Hancock S. I. , Bradley J. M. , Rumbough R. , Mcpherson J. T. , MacCormack J. N. 
Published: December 2019

ABSTRACTBoth dogs and humans can be coinfected with variousEhrlichia,Bartonella,Rickettsia, andBabesiaspecies. We investigated a kennel of sick Walker Hounds and their owners in southeastern North Carolina for evidence of tick-borne infections and associated risk factors. A high degree of coinfection was documented in the dog population. Of the 27 dogs, 26 were seroreactive to anEhrlichiasp., 16 toBabesia canis, and 25 toBartonella vinsonii, and 22 seroconverted toRickettsia rickettsiiantigens. According to PCR results, 15 dogs were infected withEhrlichia canis, 9 withEhrlichia chaffeensis, 8 withEhrlichia ewingii, 3 withEhrlichia equi, 9 withEhrlichia platys, 20 with aRickettsiaspecies, 16 with aBartonellaspecies, and 7 withB. canis. The detection of DNA from anyEhrlichiaspecies was associated with clinical illness and with concurrentB. canisinfection (by PCR). BothE. canisand an uncharacterizedRickettsiaspecies appeared to result in chronic or recurrent infection. Death in the dog population was associated with living in a dirt lot rather than the concrete kennel. Of 23 people on whom serologic testing was conducted, eight were seroreactive toBartonella henselae, one toE. chaffeensis, and one toR. rickettsiiantigen; however, none had clinical or hematologic abnormalities consistent with illness caused by these organisms. We conclude that kennel dogs with heavy tick exposure can be infected at a high rate with multiple, potentially zoonotic, tick-borne pathogens. In addition, our findings further illustrate the utility of PCR for documenting coinfection with tick-transmitted pathogens.
Evaluation of the Granada Agar Plate for Detection of Vaginal and Rectal Group B Streptococci in Pregnant Women
Authors: Gil E. García , Rodríguez M. C. , Bartolomé R. , Berjano B. , Cabero L. , Andreu A. 
Published: December 2019

ABSTRACT

Granada medium was evaluated for the detection of group B streptococci (GBS) in vaginal and rectal swabs compared with selective Columbia blood agar and selective Lim broth. From May 1996 to March 1998, 702 pregnant women (35 to 37 weeks of gestation) participated in this three-phase study; 103 (14.7%) of these women carried GBS. In the first phase of the experiment (
n
= 273 women), vaginorectal specimens were collected on the same swab; the sensitivities of Granada tube, selective Columbia blood agar, and Lim broth were 31.4, 94.3, and 74.3%, respectively. In the second and third phases (
n
= 429 women), vaginal and rectal specimens were collected separately; the sensitivities of Granada plate, selective Columbia blood agar, and Lim broth (subcultured at 4 h on selective Columbia agar in the second phase and at 18 to 24 h in Granada plate in the third phase) were 91.1, 83.9, and 75%, respectively, in the second phase and 88.5, 90.4, and 63.5%, respectively, in the third phase. There were no statistically significant differences in GBS recovery between the Granada agar plate and selective Columbia blood agar, but the Granada plate provided a clear advantage; the characteristic red-orange colonies produced overnight by GBS can be identified by the naked eye and is so specific that further identification is unnecessary. The use of the Granada tube and Lim broth did not result in increased isolation of GBS. In conclusion, the Granada agar plate is highly sensitive for detecting GBS in vaginal and rectal swabs from pregnant women and can provide results in 18 to 24 h.
Single Clonal Origin of a High Proportion of
Legionella pneumophila
Serogroup 1 Isolates from Patients and the Environment in the Area of Paris, France, over a 10-Year Period
Authors: Lawrence Christine , Reyrolle Monique , Dubrou Sylvie , Forey Françoise , Decludt Bénedicte , Goulvestre Claire , Matsiota-Bernard Peggy , Etienne Jerome , Nauciel Charles 
Published: December 2019

ABSTRACT

Arbitrarily primed PCR with three primers and pulsed-field gel electrophoresis were used to characterize a set of 75 clinical
Legionella pneumophila
serogroup 1 isolates, with no apparent epidemiological link, obtained from 24 hospitals in Paris, France, from 1987 to 1997. Unexpectedly, 25 clinical isolates from 15 hospitals had an identical profile (termed type A) by both methods. The same profile was subsequently found in 16 of 64 randomly selected environmental
L. pneumophila
serogroup 1 isolates from 15 different sites in the Paris area. There was no evidence of geographic clustering or a peak incidence of type A isolation. Type A has not been found in France outside the Paris area, suggesting that a particular type of
L. pneumophila
serogroup 1 is specifically present in the Paris water distribution network.
Novel Immunoblot Assay Using Four Recombinant Antigens for Diagnosis of Epstein-Barr Virus Primary Infection and Reactivation
Authors: Buisson M. , Fleurent B. , Mak M. , Morand P. , Chan L. , Ng A. , Guan M. , Chin D. , Seigneurin J. M. 
Published: December 2019

ABSTRACT
A new immunoblot assay, composed of four Epstein-Barr virus (EBV)-encoded recombinant proteins (virus capsid antigen [VCA] p23, early antigen [EA] p138, EA p54, and EBNA-1 p72), was compared with an immunofluorescence assay on a total of 291 sera. The test was accurate in 94.5% of cases of primary EBV infection, while an immunoglobulin G anti-VCA p23 band with strong intensity correlated with reactivation.
A Multidrug-Resistant Tuberculosis Microepidemic Caused by Genetically Closely Related
Mycobacterium tuberculosis
Strains
Authors: Kubín M. , Havelková M. , Hynčicová I. , Švecová Z. , Kaustová J. , Kremer K. , van Soolingen D. 
Published: December 2019

ABSTRACT

IS
6110
DNA fingerprinting was used to characterize an outbreak of multidrug-resistant tuberculosis in 21 individuals (17 males and 4 females) living in or roaming among four distantly separated areas in the Czech Republic. The restriction fragment length polymorphism (RFLP) analysis separated the collected
Mycobacterium tuberculosis
strains into group A, including 14 patients with six IS
6110
copies, and group B, with 7 patients displaying highly similar RFLP patterns but with two additional IS
6110
bands. A switch from pattern A to pattern B was observed in one patient, and the subsequent detection of subclone B in seven more individuals has been explained by the instability of DNA genotypes caused by transposition of IS
6110
elements.
Typing of Clinical Herpes Simplex Virus Type 1 and Type 2 Isolates with Monoclonal Antibodies
Authors: Liljeqvist Jan-Åke , Svennerholm Bo , Bergström Tomas 
Published: December 2019

ABSTRACT
The purpose of this study was to evaluate the performance of a herpes simplex virus (HSV) type 1-specific anti-glycoprotein C-1 monoclonal antibody (MAb) and a type 2-specific anti-glycoprotein G-2 MAb for typing of 2,400 clinical HSV-1 isolates and 2,400 clinical HSV-2 isolates, respectively, using an enzyme immunoassay. The anti-HSV-1 MAb showed sensitivity and specificity of 100%, and the anti-HSV-2 MAb showed a sensitivity of 99.46% and 100% specificity, indicating that these MAbs are suitable for typing of clinical HSV isolates.
Sequencing of the Ribosomal Intergenic Spacer Region for Strain Identification of
Porphyromonas gingivalis
Authors: Rumpf Robert W. , Griffen Ann L. , Wen Bo-Gui , Leys Eugene J. 
Published: December 2019

ABSTRACT

The ribosomal intergenic spacer regions (ISRs) of 19 laboratory strains and 30 clinical samples of
Porphyromonas gingivalis
were amplified by PCR and sequenced to provide a strain identifier. The ISR is a variable region of DNA located between the conserved 16S and 23S rRNA genes. This makes it an ideal locus for differentiation of strains within a species: primers specific for the conserved flanking genes were used to amplify the ISR, which was then sequenced to identify the strain. We have constructed a
P. gingivalis
ISR sequence database to facilitate strain identification. ISR sequence analysis provides a strain identifier that can be easily reproduced among laboratories and catalogued for unambiguous comparison.
Fluorescence In Situ Hybridization Assay Using Peptide Nucleic Acid Probes for Differentiation between Tuberculous and Nontuberculous Mycobacterium Species in Smears of Mycobacterium Cultures
Authors: Stender Henrik , Lund Kaare , Petersen Kenneth H. , Rasmussen Ole F. , Hongmanee Poonpilas , Miörner Håkan , Godtfredsen Sven E. 
Published: December 2019

ABSTRACT

TB PNA FISH is a new fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for differentiation between species of the
Mycobacterium tuberculosis
complex (MTC) and nontuberculous mycobacteria (NTM) in acid-fast bacillus-positive (AFB+) cultures is described. The test is based on fluorescein-labelled PNA probes that target the rRNA of MTC or NTM species applied to smears of AFB+ cultures for microscopic examination. Parallel testing with the two probes serves as an internal control for each sample such that a valid test result is based on one positive and one negative reaction. TB PNA FISH was evaluated with 30 AFB+ cultures from Denmark and 42 AFB+ cultures from Thailand. The MTC-specific PNA probe showed diagnostic sensitivities of 84 and 97%, respectively, and a diagnostic specificity of 100% in both studies, whereas the NTM-specific PNA probe showed diagnostic sensitivities of 91 and 64%, respectively, and a diagnostic specificity of 100% in both studies. The low sensitivity of the NTM-specific PNA probe in the Thai study was due to a relatively high prevalence of
Mycobacterium fortuitum
, which is not identified by the probe. In total, 63 (87%) of the cultures were correctly identified as MTC (
n
= 46) or NTM (
n
= 17), whereas the remaining 9 were negative with both probes and thus the results were inconclusive. None of the samples were incorrectly identified as MTC or NTM; thus, the predictive value of a valid test result obtained with TB PNA FISH was 100%.
Rapid Detection of Human Rhinoviruses in Nasopharyngeal Aspirates by a Microwell Reverse Transcription-PCR–Hybridization Assay
Authors: Blomqvist S. , Skyttä A. , Roivainen M. , Hovi T. 
Published: December 2019

ABSTRACT
A rapid and sensitive microwell reverse transcription (RT)-PCR–hybridization assay was developed to detect human rhinoviruses in clinical specimens and cell culture suspensions. Two hundred three nasopharyngeal aspirates collected from children with symptoms of respiratory disease were analyzed by a classical rolling-tube cell culture method, microwell culture of HeLa Ohio cell monolayers, and RT-PCR with detection of the amplicons in a microwell hybridization assay. The RT-PCR was also done with harvests of the microwell cultures. RNA was extracted with a commercial kit, and the RT-PCR procedure was carried out with microtiter-format equipment. A confirmatory test that exploited a blocking oligonucleotide at the hybridization step was developed to reliably identify marginally positive specimens. Of the 203 nasopharyngeal aspirate specimens, rhinovirus or rhinoviral RNA was detected in 111 specimens (55%). Ninety-eight specimens (48%) were found to be positive by RT-PCR of the original nasopharyngeal aspirates, while the conventional rolling-tube cell culture method yielded 52 (26%) positive specimens. This RT-PCR method with solid-phase hybridization is easy to perform, sensitive, and specific and will be especially useful for analysis of large numbers of clinical specimens.
Elucidating the Origins of Nosocomial Infections with
Candida albicans
by DNA Fingerprinting with the Complex Probe Ca3
Authors: Marco F. , Lockhart S. R. , Pfaller M. A. , Pujol C. , Rangel-Frausto M. S. , Wiblin T. , Blumberg H. M. , Edwards J. E. , Jarvis W. , Saiman L. , Patterson J. E. , Rinaldi M. G. , Wenzel R. P. ,   The Nemis Study Group, Soll D. R. 
Published: December 2019

ABSTRACT

Computer-assisted DNA fingerprinting with the complex probe Ca3 has been used to analyze the relatedness of isolates collected from individuals with nosocomial bloodstream infections (BSIs) and hospital care workers (HCWs) in the surgical and neonatal intensive care units (ICUs) of four hospitals. The results demonstrate that for the majority of patients (90%), isolates collected from commensal sites before and after collection of a BSI isolate were highly similar or identical to the BSI isolate. In addition, the average similarity coefficient for BSI isolates was similar to that for unrelated control isolates. However, the cluster characteristics of BSI isolates in dendrograms generated for each hospital compared to those of unrelated control isolates in a dendrogram demonstrated a higher degree of clustering of the former. In addition, a higher degree of clustering was observed in mixed dendrograms for HCV isolates and BSI isolates for each of the four test hospitals. In most cases, HCW isolates from an ICU were collected after the related BSI isolate, but in a few cases, the reverse was true. Although the results demonstrate that single, dominant endemic strains are not responsible for nosocomial BSIs in neonatal ICUs and surgical ICUs, they suggest that multiple endemic strains may be responsible for a significant number of cases. The results also suggest that cross-contamination occurs between patients and HCWs and between HCWs in the same ICU and in different ICUs. The temporal sequence of isolation also suggests that in the majority of cases HCWs are contaminated by isolates from colonized patients, but in a significant minority, the reverse is true. The results of this study provide the framework for a strategy for more definitive testing of the origins of
Candida albicans
strains responsible for nosocomial infections.
Epidemiology of Feline Foamy Virus and Feline Immunodeficiency Virus Infections in Domestic and Feral Cats: a Seroepidemiological Study
Authors: Winkler I. G. , Löchelt M. , Flower R. L. P. 
Published: December 2019

ABSTRACT
Although foamy viruses (Spumaviruses) have repeatedly been isolated from both healthy and diseased cats, cattle, and primates, the primary mode of transmission of those common viruses remains undefined. A database of the feline foamy virus (FeFV) and feline immunodeficiency virus (FIV) antibody status, age, and sex of 389 domestic cats presented to veterinarians was assembled. A similar database for 66 feral (wild) cats was also assembled. That FeFV antibody status reflects infection was validated by PCR. Both FeFV and FIV infection rates were found to gradually increase with age, and over 70% of cats older than 9 years were seropositive for FeFV. In domestic cats, the prevalence of FeFV infection was similar in both sexes. In feral cats, FeFV infection was more prevalent in female cats than in male cats. Although both FeFV and FIV have been reported to be transmitted by biting, the patterns of infection observed are more consistent with an interpretation that transmission of these two retroviruses is not the same. The prevalence of FIV infection is highest in nondesexed male cats, the animals most likely to display aggressive behavior. The gradual increase in the proportion of FeFV-infected animals is consistent with transmission of foamy viruses by intimate social contact between animals and less commonly by aggressive behavior.
Monitoring of Epstein-Barr Virus DNA Load in Peripheral Blood by Quantitative Competitive PCR
Authors: Stevens Servi J. C. , Vervoort Marcel B. H. J. , van den Brule Adriaan J. C. , Meenhorst Pieter L. , Meijer Chris J. L. M. , Middeldorp Jaap M. 
Published: December 2019

ABSTRACT
A competitive quantitative PCR (Q-PCR) assay combined with simple silica-based DNA extraction was developed for monitoring of Epstein-Barr virus (EBV) DNA load in unfractionated peripheral blood. The Q-PCR is based on competitive coamplification of a highly conserved 213-bp region of the EBNA-1 open reading frame with an internal standard (IS), added in a known concentration. The IS has the same amplicon length and base composition as the wild-type (WT) EBNA-1 amplicon but differs in 23 internally randomized bases. Competitive coamplification yields two PCR products that are quantified by enzyme immunoassay or by electrochemiluminescence detection, with probes specific for the 23 differing internal nucleotides. The Q-PCR has a sensitivity of 10 copies of either WT or IS plasmid DNA. The Q-PCR was validated by quantification of known amounts of plasmid containing the WT EBNA-1 target. Furthermore, we determined EBV genome copy numbers in different cell lines. For EBV quantification in clinical samples, DNA was isolated from lysed whole blood by silica-affinity purification. Forty-six percent of healthy donor peripheral blood samples were positive by Q-PCR. In most of these samples, viral load was less than 2,000 EBV copies/ml of blood. In peripheral blood samples from two AIDS-related non-Hodgkin’s lymphoma patients, elevated EBV loads (up to 120,000 copies/ml) were observed, which decreased upon therapy. In Burkitt’s lymphoma patients, up to 4,592,000 EBV genome copies/ml of blood were detected. In conclusion, the EBNA-1-based Q-PCR assay provides a reproducible, accurate, and easy method for studying the relationship between EBV load and clinical parameters.
Detection of
Leishmania infantum
in Dogs by PCR with Lymph Node Aspirates and Blood
Authors: Reale S. , Maxia L. , Vitale F. , Glorioso N. S. , Caracappa S. , Vesco G. 
Published: December 2019

ABSTRACT

The PCR technique was applied to the diagnosis of leishmaniasis in dogs, both serologically negative and positive. DNA was taken from lymph node aspirates and blood. The primers 13a and 13b, derived from
Leishmania amazonies
and
Leishmania braziliensis
kinetoplast DNA (kDNA), also amplified
Leishmania infantum
IPT1 constant region of minicircle kDNA. The amplified fragment is 116 bp long. It was cloned and the sequence was determined. A 70-bp inner fragment was designed and used as a probe in dot blot hybridization. A group of 124 dogs was examined, 37 of which showed typical symptoms of disease. PCR was performed on 124 blood samples and 52 lymph node aspirates. Using microscopic examination as the “gold standard,” we calculated sensitivity, specificity, and positive and negative predictive values of 100% using lymph node aspirates and values of 85, 80, 95, and 57%, respectively, using blood samples. We found that 40% of the animals without lesions and 38% of the animals with clinical signs gave false-negative results by indirect immunofluorescence antibody testing. These animals could contribute to the spreading of infection among dogs, and represent a potential risk for human health as well.
Comparison of Susceptibility Testing Methods with
mecA
Gene Analysis for Determining Oxacillin (Methicillin) Resistance in Clinical Isolates of
Staphylococcus aureus
and Coagulase-Negative
Staphylococcus
spp
Authors: Kohner P. , Uhl J. , Kolbert C. , Persing D. , Cockerill F. 
Published: December 2019

ABSTRACT

Ninety-nine clinical staphylococcal isolates (58 coagulase-negative
Staphylococcus
spp. [CoNS] and 41
Staphylococcus aureus
isolates) were evaluated for susceptibility to oxacillin. The following susceptibility testing methods, media, and incubation conditions were studied: agar dilution by using Mueller-Hinton (MH) medium (Difco) supplemented with either 0, 2, or 4% NaCl and incubation at 30 or 35°C in ambient air for 24 or 48 h; disk diffusion by using commercially prepared MH medium (Difco) and MH II agar (BBL) and incubation at 35°C in ambient air for 24 or 48 h; and agar screen (spot or swab inoculation) by using commercially prepared agar (Remel) or MH agar (Difco) prepared in-house, each containing 4% NaCl and 6 μg of oxacillin/ml (0.6-μg/ml oxacillin was also studied with MH agar prepared in-house for the agar swab method and CoNS isolates) and incubation at 35°C in ambient air for 24 or 48 h for swab inoculation and at 30 or 35°C in ambient air for 24 or 48 h for spot inoculation. The results for these methods were compared to the results for
mecA
gene detection by a PCR method. Given the ability to support growth and the results for susceptibility testing (the breakpoint for susceptible isolates was ≤2 μg/ml), the best methods for CoNS isolates were (i) agar dilution by using MH medium supplemented with 4% NaCl and incubation at 35°C for 48 h (no growth failures were noted, and sensitivity was 97.6%) and (ii) agar screen (swab inoculation) by using MH medium prepared in-house supplemented with 4% NaCl and containing 0.6 μg oxacillin/ml and incubation at 35°C for 48 h (one isolate that did not carry the
mecA
gene did not grow, and the sensitivity was 100%). All but one (agar dilution without added NaCl and incubation at 30°C for 48 h) of the methods tested revealed all oxacillin-resistant
S. aureus
isolates, and no growth failures occurred with any method. If the breakpoint for susceptibility was lowered to ≤1 μg/ml for agar dilution methods, more CoNS isolates with oxacillin resistance related to the
mecA
gene were detected when 0 or 2% NaCl agar supplementation was used. Only one CoNS isolate with
mecA
gene-associated resistance was not detected by using agar dilution and MH medium supplemented with 4% NaCl with incubation for 48 h. When the breakpoint for susceptibility was decreased 10-fold (from 6.0 to 0.6 μg of oxacillin per ml) for the agar swab screen method, fully 100% of the CoNS isolates that carried the
mecA
gene were identified.