bluuebunny/crossref_metadata_2025_split huggingface dataset:
Serologic Diagnosis of Lyme Borreliosis by Using Enzyme-Linked Immunosorbent Assays with Recombinant Antigens
Authors: Magnarelli Louis A. , Ijdo Jacob W. , Padula Steven J. , Flavell Richard A. , Fikrig Erol 
Published: December 2019

ABSTRACT

Class-specific enzyme-linked immunosorbent assays (ELISAs) with purified recombinant antigens of
Borrelia burgdorferi
sensu stricto and Western blot analyses with whole cells of this spirochete were used to test human sera to determine which antigens were diagnostically important. In analyses for immunoglobulin M (IgM) antibodies, 14 (82%) of 17 serum samples from persons who had erythema migrans reacted positively by an ELISA with one or more recombinant antigens. There was frequent antibody reactivity to protein 41-G (p41-G), outer surface protein C (OspC), and OspF antigens. In an ELISA for IgG antibodies, 13 (87%) of 15 serum samples had antibodies to recombinant antigens; reactivity to p22, p39, p41-G, OspC, and OspF antigens was frequent. By both ELISAs, serum specimens positive for OspB, OspE, and p37 were uncommon. Analyses of sera obtained from persons who were suspected of having human granulocytic ehrlichiosis (HGE) but who lacked antibodies to ehrlichiae revealed IgM antibodies to all recombinant antigens of
B. burgdorferi
except OspB and IgG antibodies to all antigens except OspE. Immunoblotting of sera from the study group of individuals suspected of having HGE reaffirmed antibody reactivity to multiple antigens of
B. burgdorferi
. There was minor cross-reactivity when sera from healthy subjects or persons who had syphilis, oral infections, or rheumatoid arthritis were tested by ELISAs with p37, p41-G, OspB, OspC, OspE, and OspF antigens. Although the results of class-specific ELISAs with recombinant antigens were comparable to those recorded for assays with whole-cell antigen and for individuals with confirmed clinical diagnoses of Lyme borreliosis, immunoblotting is still advised as an adjunct procedure, particularly when there are low antibody titers by an ELISA.
Detection of Anti-Arboviral Immunoglobulin G by Using a Monoclonal Antibody-Based Capture Enzyme-Linked Immunosorbent Assay
Authors: Johnson Alison J. , Martin Denise A. , Karabatsos Nick , Roehrig John T. 
Published: December 2019

ABSTRACTMonoclonal antibody (MAb)-based capture enzyme-linked immunosorbent assays (ELISAs) for the detection of anti-arboviral immunoglobulin G (IgG ELISAs) were developed for a comprehensive array of medically important arboviruses from theAlphavirus,Flavivirus, andBunyavirusgenera. Tests were optimized and standardized so that maximum homology could be maintained among working parameters for the different viral agents, enabling a wide range of viruses to be easily tested for at one time. MAbs were screened for suitability as capture vehicles for antigens from the three genera. The final test configuration utilized group-reactive MAbs eastern equine encephalitis virus 1A4B-6, dengue 2 virus 4G2, and La Crosse encephalitis virus 10G5.4 to capture the specific inactivated viral antigens. Serum IgG was detected by using alkaline phosphatase-conjugated anti-human IgG (Fc portion). A dilution of 1:400 was chosen as the universal screening serum dilution, with endpoint titrations of serum samples testing positive eliminating occasional false-positive results. IgG ELISA results correlated with those of the standard plaque-reduction neutralization assays. As expected, some test cross-reactivity was encountered within the individual genera, and tests were interpreted within the context of these reactions. The tests were standardized for laboratory diagnosis of arboviral infections, with the intent that they be used in tandem with the corresponding IgM antibody-capture ELISAs.
Transfer of Erythromycin Resistance from Poultry to Human Clinical Strains ofStaphylococcus aureus
Authors: Khan Saeed A. , Nawaz Mohamed S. , Khan Ashraf A. , Cerniglia Carl E. 
Published: December 2019

ABSTRACTThe transfer ofermAandermCgenes, the two most common resistance determinants of erythromycin resistance, was studied with Luria-Bertani broth in the absence of additional Ca2+or Mg2+ions. Fifteen human and five poultry isolates ofStaphylococcus aureus, which were resistant to erythromycin but carried different genetic markers for erythromycin resistance, were used for conjugation. Since both the donors (Amps-Tetr) and recipients (Ampr-Tets) were resistant to erythromycin, the transconjugants were initially picked up as ampicillin- and tetracycline-resistant colonies. The resistance transfer mechanisms of the chromosomally located erythromycin rRNA methylase geneermAand the plasmid-borneermCgene were monitored by a multiplex PCR and gene-specific internal probing assay. Four groups of transconjugants, based upon the transfer of theermAand/orermCgene, were distinguished from each other by the use of this method. Selective antibiotic screening revealed only one type of transconjugant that was resistant to ampicillin and tetracycline. A high frequency of transfer (4.5 × 10−3) was observed in all of the 23 transconjugants obtained, and the direction of tetracycline and erythromycin resistance marker transfer was determined to be from poultry to clinical isolates. The transfers of theermAandermCgenes were via transposition and transformation, respectively.
Genetic Relatedness of
Salmonella
Isolates from Nondomestic Birds in Southeastern United States
Authors: Hudson Charlene R. , Quist Charlotte , Lee Margie D. , Keyes Kathleen , Dodson Sara V. , Morales Cesar , Sanchez Susan , White David G. , Maurer John J. 
Published: December 2019

ABSTRACT

Salmonella
infections have been implicated in large-scale die-offs of wild birds in the United States. Although we know quite a bit about the epidemiology of
Salmonella
infection among domestic fowl, we know little about the incidence, epidemiology, and genetic relatedness of salmonellae in nondomestic birds. To gain further insight into salmonellae in these hosts, 22
Salmonella
isolates from diseased nondomestic birds were screened for the presence of virulence and antibiotic resistance-associated genes and compared genetically using pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA analysis. Of the 22
Salmonella
isolates examined, 15 were positive for the invasion gene
invA
and the virulence plasmid-associated genes
spvC
and
pef
. Most (15 of 22) were generally sensitive to antibiotics. However, two
Salmonella
isolates from pet birds were identified as
Salmonella enterica
serovar Typhimurium DT104. Despite the general susceptibility of these
Salmonella
isolates to most antimicrobial agents, antibiotic resistance-associated genes
intI1
,
merA
, and
aadA1
were identified in a number of these isolates. Five distinct
Xba
I and nine distinct
Bln
I DNA patterns were observed for the 22
Salmonella
isolates typed by PFGE. PFGE analysis determined that
Salmonella
isolates from passerines in Georgia and Wyoming were genetically related.
Bilateral Pulmonary Aspergilloma Caused by an Atypical Isolate of
Aspergillus terreus
Authors: Khan Z. U. , Kortom M. , Marouf R. , Chandy R. , Rinaldi M. G. , Sutton D. A. 
Published: December 2019

ABSTRACT

A case of bilateral pulmonary aspergilloma caused by an atypical isolate of
Aspergillus terreus
is described. The diagnosis was established by the presence of septate, dichotomously branched fungal elements in freshly collected bronchoalveolar lavage and sputum specimens and by repeated isolation of the fungus in culture. Specific precipitating antibodies against the
A. terreus
isolate were demonstrated in the patient's serum. The isolate was atypical as it failed to produce fruiting structures on routine mycological media, but it did so on extended incubation on potato flake agar and produced globose, relatively heavy-walled, hyaline accessory conidia (formerly termed aleurioconidia) on both vegetative and aerial mycelia. Also, it produced an intense yellow diffusing pigment in the medium. The report underscores the increasing importance of
A. terreus
in the etiology of pulmonary aspergillosis. It is suggested that
A. terreus
antigen be included in the battery of serodiagnostic reagents to facilitate the early diagnosis of infections caused by this species.
Isolation of an Unusual
Mycobacterium
Species from an AIDS Patient with Acute Lymphadenitis
Authors: Bajolet Odile , Beguinot Isabelle , Brasme Lucien , Jaussaud Roland , Ingrand Didier , Vincent Véronique 
Published: December 2019

ABSTRACT

A nonchromogenic
Mycobacterium
species was isolated from an AIDS patient with acute lymphadenitis. On the basis of the results of conventional tests, the strain appeared to be an atypical nonphotochromogenic
Mycobacterium kansasii
strain. Sequencing of the 16S rRNA gene revealed a unique nucleic acid sequence, suggesting that the isolate represents an undescribed pathogenic species.
A Multiplex Approach to Molecular Detection ofBrucella abortusand/orMycobacterium bovisInfection in Cattle
Authors: Sreevatsan Srinand , Bookout Jack B. , Ringpis Fidel , Perumaalla Veera S. , Ficht Thomas A. , Adams L. Garry , Hagius Sue D. , Elzer Philip H. , Bricker Betsy J. , Kumar Girish K. , Rajasekhar M. , Isloor Srikrishna , Barathur Raj R. 
Published: December 2019

ABSTRACTA multiplex amplification and detection platform for the diagnosis ofMycobacterium bovisandBrucella abortusinfection simultaneously in bovine milk and nasal secretions was developed. This system (designated the bovine pathogen detection assay [BPDA]-PCR) consists of duplex amplification of species-specific targets (a region of the BCSP31K gene ofB. abortusand a repeat-sequence region in thehsp65gene ofM. bovis, respectively). This is followed by a solid-phase probe capture hybridization of amplicons for detection. On the basis of spiking experiments with normal milk, the analytical sensitivity of the assay was 800 CFU equivalents/ml of milk forB. abortusand as low as 4 CFU equivalents per ml of milk forM. bovis. BPDA-PCR was validated with 45 liver samples from lemmings experimentally infected withB. abortus. The assay sensitivity, based on culture status as a “gold standard,” was 93.9%. In this experiment, BPDA-PCR also identified five culture-negative liver samples as positive (41.7%). Field studies for the evaluation of BPDA-PCR were performed with samples from dairy animals from geographically distinct regions (India, Mexico, and Argentina). A high prevalence of shedding ofB. abortus(samples from India) andM. bovis(samples from Mexico) was identified by BPDA-PCR. In samples from India,B. abortusshedding was identified in 86% of milk ring test-positive animals (n= 15) and 80% of milk ring test-negative cows (n= 5). In samples from Mexico,M. boviswas identified by PCR in 32.6% of pools (n= 46) of milk that each contained milk from 10 animals and in 56.2% of nasal swabs (n= 121) from cattle from tuberculin test-positive herds. In contrast, the Argentine cattle (n= 70) had a modest prevalence ofM. bovisshedding in nasal swabs (2.9%) and milk (1.4%) and ofB. abortusin milk (11.4%). On the basis of these analyses, we identify BPDA-PCR as an optimal tool for both screening of herds and testing of individual animals in a disease eradication program. A combination of the duplex assay, screening of milk samples in pools, and the proposed algorithm provides a highly sensitive, cost-effective, and economically viable alternative to serological testing.
No Evidence of Measles Virus in Stapes Samples from Patients with Otosclerosis
Authors: Grayeli Alexis Bozorg , Palmer Pierre , Tran Ba Huy Patrice , Soudant Jacques , Sterkers Olivier , Lebon Pierre , Ferrary Evelyne 
Published: December 2019

ABSTRACT

Otosclerosis is a localized bone dystrophy of unknown etiology mainly involving the stapes. The hypothesis of a persistent infection by the measles virus was based on the inconstant detection of the virus by various methods, including reverse transcription-PCR (RT-PCR) of patients' stapes samples. The aim of this work was to investigate the presence of the measles virus in stapedial otosclerosis foci by different sensitive methods. Pathologic stapes samples were obtained from 35 patients suffering from otosclerosis. Measles virus detection was performed by (i) cocultures of Vero cells and primary cell cultures of bone samples (
n
= 7), (ii) immunofluorescence study of these cocultures (
n
= 3), and (iii) RT-PCR on RNA directly obtained from fresh frozen samples (
n
= 28) and on RNA extracted from the primary cell cultures (
n
= 2). Viral genomic regions coding for N (nucleoprotein) and M (matrix) proteins were separately amplified. PCR sensitivity was optimized on the measles virus Edmonston strain. Glyceraldehyde-3-phosphate dehydrogenase mRNA was used as a marker of total RNA recovery. PCR products were tested by Southern blot hybridization technique to improve sensitivity and specificity. PCRs amplifying the M and the N protein genes were able to detect the control measles virus RNA at titers as low as 0.1 and 0.01 50% tissue culture infective dose, respectively. With these highly sensitive methods, we could not evidence the presence of the measles virus in any of our bone samples or primary bone cell cultures. Our results do not confirm the hypothesis of persistent measles virus infection in otosclerosis.
Evaluation of Performance of the Gen-Probe Human Immunodeficiency Virus Type 1 Viral Load Assay Using Primary Subtype A, C, and D Isolates from Kenya
Authors: Emery Sandra , Bodrug Sharon , Richardson Barbra A. , Giachetti Cristina , Bott Martha A. , Panteleeff Dana , Jagodzinski Linda L. , Michael Nelson L. , Nduati Ruth , Bwayo Job , Kreiss Joan K. , Overbaugh Julie 
Published: December 2019

ABSTRACT

Accurate and sensitive quantification of human immunodeficiency virus type 1 (HIV-1) RNA has been invaluable as a marker for disease prognosis and for clinical monitoring of HIV-1 disease. The first generation of commercially available HIV-1 RNA tests were optimized to detect the predominant HIV-1 subtype found in North America and Europe, subtype B. However, these tests are frequently suboptimal in detecting HIV-1 genetic forms or subtypes found in other parts of the world. The goal of the present study was to evaluate the performance of a new viral load assay with non-subtype B viruses. A transcription-mediated amplification method for detection and quantitation of diverse HIV-1 subtypes, called the Gen-Probe HIV-1 viral load assay, is under development. In this study we examined the performance of the Gen-Probe HIV-1 viral load assay relative to that of the commonly used commercial HIV-1 RNA assays using a panel of primary isolates from Kenya. For comparison, we included several subtype B cloned viruses, and we quantified each virus using an in-house quantitative-competitive reverse transcriptase PCR (QC-RT-PCR) method and gag
p24
antigen capture. The Gen-Probe HIV-1 viral load assay and a version of the Roche AMPLICOR HIV-1 MONITOR test (version 1.5) that was designed to detect a broader range of subtypes were both sensitive for the quantification of Kenyan primary isolates, which represented subtype A, C, and D viruses. The Gen-Probe HIV-1 viral load assay was more sensitive for the majority of viruses than the Roche AMPLICOR HIV-1 MONITOR test version 1.0, the Bayer Quantiplex HIV RNA 3.0 assay, or a QC-RT-PCR method in use in our laboratory, suggesting that it provides a useful method for quantifying HIV-1 RNAs from diverse parts of the world, including Africa.
Detection ofLeptospiraspp. in the Aqueous Humor of Horses with Naturally Acquired Recurrent Uveitis
Authors: Faber Nick A. , Crawford Melissa , LeFebvre Rance B. , Buyukmihci Nedim C. , Madigan John E. , Willits Neil H. 
Published: December 2019

ABSTRACTLeptospiral organisms have long been presumed to be associated with the presence of equine recurrent uveitis. This project was undertaken to determine the presence ofLeptospiraspp. in the aqueous humor of horses with uveitis to determine if there was an association with inflammation. Thirty horses were determined to have recurrent uveitis based on clinical evaluation or history. Sixteen horses were judged clinically and historically to be free of uveitis and were used as controls. Aqueous humor samples were cultured and evaluated by PCR for the presence ofLeptospiraDNA. Serum was collected and evaluated for the presence of antibodies against five serovars in a leptospirosis panel. Twenty-one of 30 horses with recurrent uveitis and one of 16 uveitis-free horses were positive by PCR for the presence ofLeptospiraDNA. Six of these 21 horses with uveitis were culture positive for leptospires from the aqueous humor. Serologic results did not correlate well with the presence ofLeptospiraDNA or organisms in the aqueous humor.Leptospiraspp. are present in a high percentage of horses with naturally occurring recurrent uveitis.
Identification of a Unique
Helicobacter
Species by 16S rRNA Gene Analysis in an Abdominal Abscess from a Patient with X-Linked Hypogammaglobulinemia
Authors: Han S.-R. , Schindel C. , Genitsariotis R. , Märker-Hermann E. , Bhakdi S. , Maeurer M. J. 
Published: December 2019

ABSTRACT

A unique
Helicobacter
species, MZ640285, was isolated from a patient with X-linked hypogammaglobulinemia suffering from recurrent abdominal abscesses and was identified by 16S rRNA gene sequence analysis. In the phylogenetic tree, the isolate fell into a cluster which included
Flexispira rappini
,
Helicobacter bilis
, and
Helicobacter
sp. strain Mainz. Helicobacters are being increasingly recognized as pathogens in immunocompromised hosts. These fastidious bacteria are not easily cultured in the routine diagnostic laboratory, and this is the first report of their identification by 16S rRNA gene sequencing performed directly from a clinical specimen.
Survey for the Presence and Distribution of Human Herpesvirus 8 in Healthy Brain
Authors: Chan Paul K. S. , Ng Ho-Keung , Cheung Jo L. K. , Cheng Augustine F. 
Published: December 2019

ABSTRACT

Brain tissues from both sides of the cerebellum and frontal, temporal, parietal, and occipital lobes were collected postmortem from 30 patients for human herpesvirus 8 (HHV-8) detection by PCR. Overall, 42 of 300 (14.0%) samples were positive, with similar rates for each position. Nineteen patients (63.3%) were positive and showed a significant increase in positivity with age (
P
= 0.007). The results indicate a neuroinvasive and neuropersistent potential of HHV-8.
vacA
Genotypes in
Helicobacter pylori
Strains Isolated from Children with and without Duodenal Ulcer in Brazil
Authors: De Gusmão Valquiria Ribeiro , Nogueira Mendes Edilberto , De Magalhães Queiroz Dulciene Maria , Aguiar Rocha Gifone , Camargos Rocha Andreia Maria , Ramadan Ashour Abdussalam Ali , Teles Carvalho Anfrisina Sales 
Published: December 2019

ABSTRACT

Data concerning the association between
vacA
genotypes and disease in children in both developed and developing countries are scarce, especially because of the small number of children with a duodenal ulcer studied. The
vacA
genotypes of
Helicobacter pylori
strains obtained from 65 children (24 with a duodenal ulcer and 41 without a duodenal ulcer; 33 girls; mean age, 10.2 years; age range, 1 to 17 years) were investigated as described by J. C. Atherton et al. (J. Clin. Microbiol. 37:2979–2982, 1999). Ten (15.4%) children were infected with more than one
H. pylori
strain. None of these patients were included in our analysis of the relationship between gastric disorders and specific
vacA
genotypes. The s1 allele was detected in all
H. pylori
strains isolated from patients with a duodenal ulcer and from 21 (58.3%) patients without a duodenal ulcer (
P
= 0.003). Strains with the s2 allele were found only in patients without ulcer (
n
= 15; 41.7%). Most s1 strains had the s1b allele (97.5%), a result similar to that reported for adults from the Iberian peninsula, which could reflect the Brazilian population origin. One untypeable s1 strain was isolated. The m1 allele was also more frequently found in strains obtained from duodenal ulcer patients (
P
= 0.028). The m2 allele was found in strains obtained from 20 (36.4%) children, 3 (15.8%) with an ulcer and 17 (47.2%) without an ulcer. Only one m hybrid strain (m1 and m2 hybrid) was detected. It was demonstrated for the first time that the frequencies of colonization with strains with the s1 allele (14.3% in children up to 8 years of age and 85.7% in older patients;
P
= 0.012) and of strains with the m1 allele (11.1% in patients up to the age 8 years and 88.9% in older children;
P
= 0.013) increase with age.
Mutations in the
rpoB
Gene of Multidrug-Resistant
Mycobacterium tuberculosis
Isolates from Brazil
Authors: Valim Andréia Rosane M. , Rossetti Maria Lucia R. , Ribeiro Marta O. , Zaha Arnaldo 
Published: December 2019

ABSTRACT

Mutations in a 69-bp region of the
rpoB
gene associated with rifampin resistance (Rif
r
) in 100 isolates (82 Rif
r
) from three states of Brazil were studied. Twenty-one different kinds of mutations were identified in the Rif
r
isolates, and six new alleles are described.
Expression of Foreign Proteins by Poliovirus Polyprotein Fusion: Analysis of Genetic Stability Reveals Rapid Deletions and Formation of Cardioviruslike Open Reading Frames
Authors: Mueller Steffen , Wimmer Eckard 
Published: December 2019

ABSTRACT

Using a strategy developed by R. Andino, D. Silvera, S. D. Suggett, P. I. Achacoso, C. J. Miller, D. Baltimore, and M. B. Feinberg (Science 265:1448–1451, 1994), we constructed recombinant polioviruses by fusing the open reading frame (ORF) of the green fluorescent protein gene (
gfp
) of
Aequorea victoria
or the
gag
gene (encoding p17-p24) of human immunodeficiency virus type 1 (HIV-1) to the N terminus of the poliovirus polyprotein. All poliovirus expression vectors constructed by us and those obtained from Andino et al. were found to be severely impaired in viral replication and genetically unstable. Upon replication, inserted sequences were rapidly deleted as early as the first growth cycle in HeLa cells. However, the vector viruses did not readily revert to the wild-type sequence but rather retained some of the insert plus the artificial 3C
pro
/3CD
pro
cleavage site, engineered between the heterologous sequence and the poliovirus polyprotein, to give rise to genotypes reminiscent of cardioviruses. These virus variants that carry a small leader polypeptide were now relatively stable, and they grew better than their progenitor strains. Reverse transcription followed by PCR and sequence analysis of the genomic RNAs reproducibly revealed a few preferred genotypes among the isolated deletion variants. The remaining truncated inserts were retained through subsequent passages. In the immediate vicinity of the deletion borders, we observed short direct sequence repeats that we propose are involved in aligning RNA strands for illegitimate (nonhomologous) RNA recombination during minus-strand synthesis. On the basis of our results, which are at variance with published data, the utility of poliovirus vectors to express proteins >10 kDa in size through fusion with the polyprotein needs to be reevaluated.