bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antibodies against Specific Proteins of and Immobilizing Activity against Three Strains ofBorrelia burgdorferiSensu Lato Can Be Found in Symptomatic but Not in Infected Asymptomatic Dogs
Authors: Hovius Joppe W. R. , Hovius K. Emil , Oei Anneke , Houwers Dirk J. , van Dam Alje P. 
Published: December 2019

ABSTRACTIn an area where Lyme disease is endemic in The Netherlands all dogs had positive titers by whole-cell enzyme-linked immunosorbent assay and appeared to be naturally infected byBorrelia burgdorferisensu lato. To compare the antibody responses of symptomatic dogs and asymptomatic controls, we performed Western blots and in vitro immobilization assays to study antibody-dependent bactericidal activity. Strains from three different genospecies were employed as the antigen source:B. burgdorferistrain B31,Borrelia gariniistrain A87S, andBorrelia afzeliistrain pKo. Antibodies against flagellin (p41) and p39 for three strains were found in sera from both symptomatic and asymptomatic dogs and were therefore considered to be markers of exposure. Antibodies against p56 and p30 of strain B31, against p75, p58, p50, OspC, and p<19 of strain A87S, and against p56, p54, p45, OspB, p31, p26, and p<19 of strain pKo were found significantly more frequently in sera from symptomatic dogs younger than 8 years when the first symptoms were observed than in those from age-matched controls (P< 0.01). These antibodies were not found in preclinical sera and appeared during development of disease. Antibodies against OspA of strains B31 and A87S were only seen in acute-phase and convalescent sera from three dogs that recovered from disease. Incubation with 25% normal canine serum did not result in the immobilization of strains B31 and pKo, but partial immobilization of strain A87S (61% ± 24% [standard deviation] at 5 h) occurred. Seven of 15 sera from symptomatic dogs but none of the sera from 11 asymptomatic dogs had antibody-dependent immobilizing activity against one of the strains. Consecutive sera from one of these dogs immobilized two different strains. Antibody-mediated bactericidal serum was not seen before onset of disease, was strongest in the acute phase of disease, and fluctuated during chronic disease. From seven out of eight symptomatic dogsBorreliaDNA was amplified by PCR; in three of them the bactericidal activity was directed against one of the genospecies amplified from that dog; however, four PCR-positive dogs lacked bactericidal activity. In conclusion, dogs with symptomatic canine borreliosis have more-extensive antibody reactivity againstBorrelia, as shown by both Western blotting and immobilization assays.
Quantitative Approach for the Serodiagnosis of Canine Lyme Disease by the Immunoblot Procedure
Authors: Guerra Marta A. , Walker Edward D. , Kitron Uriel 
Published: December 2019

ABSTRACTSerum samples obtained from healthy, asymptomatic dogs in areas of Wisconsin and northern Illinois where Lyme disease is endemic or nonendemic were assayed for antibodies toBorrelia burgdorferiby enzyme-linked immunosorbent assay (ELISA), and positive results were confirmed by immunoblot assay. We found that 56.9% (562 of 1,077) of the samples were positive by ELISA and 82.0% (461 of 562) were positive by immunoblotting. A logistic regression model was developed to distinguish between nonvaccinated dogs naturally infected withB. burgdorferifrom areas where the disease is endemic and dogs from areas where the disease is nonendemic that were vaccinated against Lyme disease. Of the 18 protein bands analyzed, 8 were found to be significantly different (P< 0.05) between the two groups. p93, p34, p31, and p28 occurred with increased frequency in vaccinated dogs, while p58, p37, p35, and p30 occurred more frequently in naturally infected dogs. The logistic regression equation obtained was used to determine the probability of natural infection among vaccinated dogs residing in areas where the disease is endemic. Of 125 samples, 87.2% had a very low probability of natural infection and only 2.4% were highly likely to be infected. Logistic regression is a useful method for distinguishing between vaccinated and naturally infected dogs and predicting the serological status of vaccinated dogs from areas where Lyme disease is endemic.
Slaughter Pigs Are Commonly Infected by Closely Related but Distinct Gastric Ulcerative Lesion-Inducing Gastrospirilla
Authors: Roosendaal Robert , Vos Jan H. , Roumen Thijs , van Vugt René , Cattoli Giovanni , Bart Aldert , Klaasen Henricus L. B. M. , Kuipers Ernst J. , Vandenbroucke-Grauls Christina M. J. E. , Kusters Johannes G. 
Published: December 2019

ABSTRACT

An association between (unculturable) gastrospirillum-like organisms (GLO) and ulcerative lesions in the pars oesophagea in stomachs of swine has been claimed. In dogs GLO detected by microscopy may represent several
Helicobacter
species or subspecies. Therefore we investigated which
Helicobacter
spp. are present in stomachs of swine and their possible association with ulcerative lesions of the pars oesophagea. The presence of
Helicobacter
spp. in the antrum and pars oesophagea in 122 stomachs of slaughter swine was determined by microscopy (
n
= 122), by culture on selective and nonselective media (
n
= 112), and by a genus-specific 16S ribosomal DNA (rDNA) PCR (
n
= 80). GLO could not be cultured. Phylogenetic analysis of 43 16S rDNA fragments (out of 54 PCR-positive biopsy specimens) revealed the presence of
Helicobacter heilmannii
type 1 in 42 of them. This correlated with the presence of bacteria with GLO morphology.
Helicobacter bilis
16S rDNA was amplified directly from one sample harboring bacteria with
H. bilis
morphology. The association between
Helicobacter
spp. and gastric lesions was investigated with a second group of 41 pigs with (
n
= 21 cases) or without (
n
= 20 controls) gastric lesions. Fifteen of the 21 cases were positive by PCR or microscopy, compared to 7 of 20 of the controls (
P
= 0.03). 16S rDNA sequence analysis of 7 of 14 PCR-positive cases revealed the presence of
H. heilmannii
type 1. Microscopy showed bacteria with GLO morphology. One sample (cases) was culture negative but PCR positive for
Helicobacter pullorum
-related 16S rDNA. In conclusion, our findings indicate that
H. heilmannii
type 1 is the predominant
Helicobacter
spp. in the stomachs of pigs and that its presence is associated with ulcerative lesions in the pars oesophagea.
Evaluation of a Commercially Available Recombinant-Protein Enzyme-Linked Immunosorbent Assay for Detection of Antibodies Produced in Scrub Typhus Rickettsial Infections
Authors: Land Meagan V. , Ching Wei-Mei , Dasch Gregory A. , Zhang Zhiwen , Kelly Daryl J. , Graves Stephen R. , Devine Peter L. 
Published: December 2019

ABSTRACT

The 56-kDa major outer membrane protein antigen of
Orientia tsutsugamuchi
is the immunodominant antigen in human scrub typhus (ST) infections. An enzyme-linked immunosorbent assay (ELISA) using a recombinant 56-kDa protein (r56) to detect specific immunoglobulin M (IgM) produced in ST infections was developed, and its performance was evaluated using sera from patients with active ST (n = 59), spotted fever (SF) (n = 31), and murine typhus (MT) (n = 6) and from those without rickettsial infection (n = 52). The r56 ELISA was compared to an ELISA using native whole cell lysate of
O. tsutsugamushi
Karp or
O. tsutsugamushi
Gilliam as antigens. The performance of the assays using r56 was similar to that of those using native antigens. Using indirect immunoperoxidase (IIP) as the reference test, sensitivities were 86, 88, and 88% while specificities were 84, 90, and 87% in the three assays. Furthermore, cross-reactivity in confirmed cases of SF and MT was low (5.4, 2.7, and 2.7% respectively). The additional use of IgG in the r56 ELISA gave improved performance (sensitivity, 80%; specificity, 96%; cross-reactivity in SF and MT, 2.7%). The detection of high levels of IgG in some IgM-negative patients illustrates the importance of including a test for IgG in the detection of secondary or reactivated infections, since many of these patients were from regions in Thailand where these infections are endemic.
Detection of Human Cytomegalovirus DNA by Real-Time Quantitative PCR
Authors: Nitsche Andreas , Steuer Nina , Schmidt Christian Andreas , Landt Olfert , Ellerbrok Heinz , Pauli Georg , Siegert Wolfgang 
Published: December 2019

ABSTRACT
A real-time PCR assay was developed to quantify human cytomegalovirus (CMV) DNA. This assay was used to demonstrate a higher CMV DNA load in plasma of bone marrow transplant patients than in that of blood donors. The CMV load was higher in CMV antigen-positive patients than in antigen-negative patients.
Identification of
Ehrlichia chaffeensis
by Nested PCR in Ticks from Southern China
Authors: Cao Wu-Chun , Gao Yu-Min , Zhang Pan-He , Zhang Xi-Tan , Dai Qing-Hua , Dumler J. Stephen , Fang Li-Qun , Yang Hong 
Published: December 2019

ABSTRACT

A total of 717 ticks collected from southern China were examined by nested PCR for the presence of
Ehrlichia chaffeensis
. Sixteen (55.2%) of 29 adult
Amblyomma testudinarium
ticks and 28 (11.7%) of 240 adult and at least 4.2% of 215 nymphal (pooled specimens)
Haemaphysalis yeni
ticks tested positive. Four other species of ticks were negative. Selected positive amplicons were confirmed by DNA sequencing.
Evaluation of
recA
Sequences for Identification of
Mycobacterium
Species
Authors: Blackwood Kym S. , He Cheng , Gunton James , Turenne Christine Y. , Wolfe Joyce , Kabani Amin M. 
Published: December 2019

ABSTRACT

16S rRNA sequence data have been used to provide a molecular basis for an accurate system for identification of members of the genus
Mycobacterium
. Previous studies have shown that
Mycobacterium
species demonstrate high levels (>94%) of 16S rRNA sequence similarity and that this method cannot differentiate between all species, i.e.,
M. gastri
and
M. kansasii
. In the present study, we have used the
recA
gene as an alternative sequencing target in order to complement 16S rRNA sequence-based genetic identification. The
recA
genes of 30
Mycobacterium
species were amplified by PCR, sequenced, and compared with the published
recA
sequences of
M. tuberculosis
,
M. smegmatis
, and
M. leprae
available from GenBank. By
recA
sequencing the species showed a lower degree of interspecies similarity than they did by 16S rRNA gene sequence analysis, ranging from 96.2% between
M. gastri
and
M. kansasii
to 75.7% between
M. aurum
and
M. leprae
. Exceptions to this were members of the
M. tuberculosis
complex, which were identical. Two strains of each of 27 species were tested, and the intraspecies similarity ranged from 98.7 to 100%. In addition, we identified new
Mycobacterium
species that contain a protein intron in their
recA
genes, similar to
M. tuberculosis
and
M. leprae
. We propose that
recA
gene sequencing offers a complementary method to 16S rRNA gene sequencing for the accurate identification of the
Mycobacterium
species.
PCR Fragment Length Polymorphism Analysis of Vancomycin-Resistant
Enterococcus faecium
Authors: Donabedian Susan , Hershberger Ellie , Thal Lee Ann , Chow J. W. , Clewell Don B. , Robinson-Dunn Barbara , Zervos Marcus J. 
Published: December 2019

ABSTRACT

In this study, the glycopeptide resistance element, Tn
1546
, in 124 VanA
Enterococcus faecium
clinical isolates from 13 Michigan hospitals was evaluated using PCR fragment length polymorphism. There were 26 pulsed-field gel electrophoresis (PFGE) types, which consisted of epidemiologically related and unrelated isolates from separate patients (1992 to 1996). Previously published oligonucleotides specific for regions in the
vanA
gene cluster of Tn
1546
were used to amplify
vanRS
,
vanSH
,
vanHAX
,
vanXY
, and
vanYZ
. The glycopeptide resistance element, Tn
1546
, of
E. faecium
228 was used as the basis of comparison for all the isolates in this study. Five PCR fragment length patterns were found, as follows. (i) PCR amplicons were the same size as those of EF228 for all genes in the
vanA
cluster in 19.4% of isolates. (ii) The PCR amplicon for
vanSH
was larger than that of EF228 (3.7 versus 2.3 kb) due to an insertion between the
vanS
and
vanH
genes (79.2% of isolates). (iii) One isolate in a unique PFGE group had a
vanSH
amplicon larger than that of EF228 (5.7 versus 2.3 kb) due to an insertion in the
vanS
gene and an insertion between the
vanS
and
vanH
genes. (iv) One isolate did not produce a
vanSH
amplicon, but when
vanS
and
vanH
were amplified separately, both amplicons were the same size as those as EF228. (v) One isolate had a
vanYZ
PCR product larger than that of EF228 (2.8 versus 1.6 kb). This study shows that in a majority of the VanA
E. faecium
isolates, Tn
1546
is altered compared to that of EF228. A total of 79.2% of the study isolates had the same-size insertion between the
vanS
and
vanH
genes. The results of this study show dissemination of an altered Tn
1546
in heterologous VanA
E. faecium
in Michigan hospitals.
Rapid Detection and Identification ofCandida,Aspergillus, andFusariumSpecies in Ocular Samples Using Nested PCR
Authors: Jaeger Emma E. M. , Carroll Nora M. , Choudhury Sarah , Dunlop Anthony A. S. , Towler Hamish M. A. , Matheson Melville M. , Adamson Peter , Okhravi Narciss , Lightman Susan 
Published: December 2019

ABSTRACTA protocol for the rapid detection of fungal DNA in ocular samples, derived from three species,Candida albicans,Aspergillus fumigatus, andFusarium solani, has been developed. Two novel panfungal primers complementary to 18S rRNA sequences present in all three species were designed. Panfungal PCR was followed by three nested PCRs utilizing species-specific primers. PCR sensitivity ranged from 50 to 100 fg of free DNA and between one and twoC. albicansorganisms. In addition, we also developed a rapid and reliable DNA extraction protocol. This protocol minimized DNA loss during extraction, whilst removing compounds from vitreous and aqueous fluids that have previously been shown to have inhibitory effects on PCR. Preliminary results obtained after testing the protocol on three patient samples support culture results and medical history. However, one patient was PCR positive but culture negative, suggesting that the sensitivity of this protocol may exceed that of traditional culture techniques. This system, therefore, constitutes an additional protocol that may significantly aid patient management in cases where fungal endophthalmitis is suspected.
Comparison of NCCLS and 3-(4,5-Dimethyl-2-Thiazyl)-2,5-Diphenyl-2H-Tetrazolium Bromide (MTT) Methods of In Vitro Susceptibility Testing of Filamentous Fungi and Development of a New Simplified Method
Authors: Meletiadis Joseph , Meis Jacques F. G. M. , Mouton Johan W. , Donnelly J. Peter , Verweij Paul E. 
Published: December 2019

ABSTRACT

The susceptibility of 30 clinical isolates belonging to six different species of filamentous fungi (
Aspergillus fumigatus
,
Aspergillus flavus
,
Scedosporium prolificans
,
Scedosporium apiospermum
,
Fusarium solani
, and
Fusarium oxysporum
) was tested against six antifungal drugs (miconazole, voriconazole, itraconazole, UR9825, terbinafine, and amphotericin B) with the microdilution method recommended by the National Committee for Clinical Laboratory Standards (NCCLS) (M38-P). The MICs were compared with the MICs obtained by a colorimetric method measuring the reduction of the dye 3-(4,5-dimethyl-2-thiazyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) to formazan by viable fungi. The levels of agreement between the two methods were 96 and 92% for MIC-0 (clear wells) and MIC-1 (75% growth reduction), respectively. The levels of agreement were always higher for
Aspergillus
spp. (97% ± 2.5%), followed by
Scedosporium
spp. (87% ± 10.3%) and
Fusarium
spp. (78% ± 7.8%). The NCCLS method was more reproducible than the MTT method: 98 versus 95% for MIC-0 and 97 versus 90% for MIC-1. However, the percentage of hyphal growth as determined visually by the NCCLS method showed several discrepancies when they were compared with the percentages of MTT reduction. A new simplified assay that incorporates the dye MTT with the initial inoculum and in which the fungi are incubated with the dye for 48 h or more was developed, showing comparable levels of agreement and reproducibility with the other two methods. Furthermore, the new assay was easier to perform and more sensitive than the MTT method.
Detection of Adenoviruses (AdV) in Culture-Negative Environmental Samples by PCR during an AdV-Associated Respiratory Disease Outbreak
Authors: Echavarria Marcela , Kolavic Shellie A. , Cersovsky Steven , Mitchell Felicia , Sanchez Jose L. , Polyak Christina , Innis Bruce L. , Binn Leonard N. 
Published: December 2019

ABSTRACT
Since 1954, adenoviruses (AdV) have been recognized as an important cause of acute respiratory disease (ARD) among U.S. military recruits. Until recently, routine oral vaccination for AdV serotypes 4 and 7 eliminated epidemic AdV-associated ARD in this population. Now that the manufacturer has ceased production, vaccination has ended and AdV epidemics have reappeared. As part of a prospective epidemiological study during the high-risk ARD season, serial samples were obtained from ventilation system filters and tested for AdV by culture and PCR. An outbreak occurred during this surveillance. Of 59 air filters, 26 (44%) were AdV positive only by PCR. Sequence analysis confirmed the presence of AdV serotype 4, the implicated outbreak serotype. The number of AdV-related hospitalizations was directly correlated with the proportion of filters containing AdV; correlation coefficients were 0.86 (Pearson) and 0.90 (Spearman's rho). This is the first report describing a PCR method to detect airborne AdV during an ARD outbreak. It suggests that this technique can detect and quantify AdV-associated ARD exposure and may enable further definition of environmental effects on AdV-associated ARD spread.
Outbreak of Methicillin-Resistant
Staphylococcus aureus
with Reduced Susceptibility to Glycopeptides in a Parisian Hospital
Authors: Guerin François , Buu-Hoï Annie , Mainardi Jean-Luc , Kac Guillaume , Colardelle Nathalie , Vaupré Sabine , Gutmann Laurent , Podglajen Isabelle 
Published: December 2019

ABSTRACT

Epidemiological relationships were investigated between 40 methicillin-resistant
Staphylococcus aureus
(MRSA) strains with decreased glycopeptide susceptibility isolated from November 1998 to March 1999 from 39 patients (17 infected and 22 colonized patients) in nine wards of the Broussais Hospital, Paris, France. Reduced glycopeptide susceptibility was readily detected on brain heart infusion (BHI) agar containing 6 μg of teicoplanin per ml and on gradient plates, but not by the standard disk diffusion method. The MICs of vancomycin and teicoplanin, determined on BHI agar, were 4 and 8 to 32 μg/ml, respectively (standard antibiotic dilution), and 4 to 8 and 8 to 32 μg/ml, respectively (E-test). All strains were resistant to macrolides, aminoglycosides, tetracycline, rifampin, sulfonamides, and pefloxacin, showed reduced susceptibility to fusidic acid and fosfomycin, and were susceptible to trimethoprim and chloramphenicol. Pulsed-field gel electrophoresis and lysotyping revealed that a multidrug-resistant MRSA clone with decreased susceptibility to glycopeptides has been discretely endemic since at least 1996 in our institution, where it was responsible for an outbreak in November and December 1998.
Comparison of BACTEC MYCO/F LYTIC and WAMPOLE ISOLATOR 10 (Lysis-Centrifugation) Systems for Detection of Bacteremia, Mycobacteremia, and Fungemia in a Developing Country
Authors: Archibald Lennox K. , McDonald L. Clifford , Addison Rachel M. , McKnight Celeste , Byrne Terry , Dobbie Hamish , Nwanyanwu Okey , Kazembe Peter , Reller L. Barth , Jarvis William R. 
Published: December 2019

ABSTRACT

In less-developed countries, studies of bloodstream infections (BSI) have been hindered because of the difficulty and costs of culturing blood for bacteria, mycobacteria, and fungi. During two study periods (study period I [1997] and study period II [1998]), we cultured blood from patients in Malawi by using the BACTEC MYCO/F LYTIC (MFL), ISOLATOR 10 (Isolator), Septi-Chek AFB (SC-AFB), and Septi-Chek bacterial (SC-B) systems. During study period I, blood was inoculated at 5 ml into an MFL bottle, 10 ml into an Isolator tube for lysis and centrifugation, and 10 ml into an SC-B bottle. Next, 0.5-ml aliquots of Isolator concentrate were inoculated into an SC-AFB bottle and onto Middlebrook 7H11 agar slants, chocolate agar slants, and Inhibitory Mold Agar (IMA) slants. During study period II, the SC-B and chocolate agar cultures were discontinued. MFL growth was detected by fluorescence caused by shining UV light (λ = 365 nm) onto the indicator on the bottom of the bottle. During study period I, 251 blood cultures yielded 44 bacterial isolates. For bacteremia, the MFL was similar to the Isolator concentrate on chocolate agar (34 of 44 versus 27 of 44;
P
, not significant [NS]), but more sensitive than the SC-B bottle (34 of 44 versus 24 of 44;
P
= 0.05). For both study periods combined, 486 blood cultures yielded 37 mycobacterial and 13 fungal isolates. For mycobacteremia, the sensitivities of the MFL and Isolator concentrate in the SC-AFB bottle were similar (30 of 37 versus 29 of 37;
P
, NS); the MFL bottle was more sensitive than the concentrate on Middlebrook agar (30 of 37 versus 15 of 37;
P
= 0.002). For fungemia, the MFL bottle was as sensitive as the SC-B bottle or Isolator concentrate on chocolate agar or IMA slants. We conclude that the MFL bottle, inoculated with just 5 ml of blood and examined under UV light, provides a sensitive and uncomplicated method for comprehensive detection of BSI in less-developed countries.
Simple and Rapid Detection of
Candida albicans
DNA in Serum by PCR for Diagnosis of Invasive Candidiasis
Authors: Wahyuningsih Retno , Freisleben Hans-Joachim , Sonntag Hans-Günther , Schnitzler Paul 
Published: December 2019

ABSTRACT

A rapid and sensitive PCR assay for the detection of
Candida albicans
DNA in serum was established. DNA from human serum samples was purified using the QIAamp blood kit, which proved to be a fast and simple method for isolating minute amounts of
Candida
DNA from clinical specimens for diagnosis of invasive candidiasis. Universal primer sequences used in the PCR assay are derived from the internal transcribed spacer rRNA gene of fungi, whereas the biotinylated hybridization probe used in a DNA enzyme immunoassay (DEIA) binds specifically to
C. albicans
DNA. The sensitivity of this PCR-DEIA method is very high; the detection limit for genomic
Candida
DNA is one
C. albicans
genome per assay. Blood from uninfected and infected persons, ranging from healthy volunteers, patients with mucocutaneous infections, and patients at risk to develop a systemic
Candida
infection to patients with an established systemic candidiasis, was analyzed for the presence of
C. albicans
to diagnose fungal infection.
Candida
DNA could not be detected in sera of 16 culture-negative controls and from 11 nonsystemic candidal infections by PCR or DEIA. Blood cultures from patients at risk were all negative for
Candida
, whereas all blood cultures from systemic candidiasis patients were positive. However,
Candida
DNA could be detected by PCR and DEIA in the serum from three out of nine patients who were at risk for a systemic infection and in the serum of all seven patients who had already developed an invasive
Candida
infection. PCR is more sensitive than blood culture, since some of the patients at risk for invasive yeast infection, whose blood cultures were all negative for
Candida
, tested positive in the PCR amplification. These results indicate the potential value of PCR for detecting
C. albicans
in serum samples and for identifying patients at risk for invasive candidiasis.