bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human Rhinovirus 87 and Enterovirus 68 Represent a Unique Serotype with Rhinovirus and Enterovirus Features
Authors: Blomqvist Soile , Savolainen Carita , Råman Laura , Roivainen Merja , Hovi Tapani 
Published: October 2002

ABSTRACT

It has recently been reported that all but one of the 102 known serotypes of the genus
Rhinovirus
segregate into two genetic clusters (C. Savolainen, S. Blomqvist, M. N. Mulders, and T. Hovi, J. Gen. Virol.
83:
333-340, 2002). The only exception is human rhinovirus 87 (HRV87). Here we demonstrate that HRV87 is genetically and antigenically highly similar to enterovirus 68 (EV68) and is related to EV70, the other member of human enterovirus group D. The partial nucleotide sequences of the 5′ untranslated region, capsid regions VP4/VP2 and VP1, and the 3D RNA polymerase gene of the HRV87 prototype strain F02-3607 Corn showed 97.3, 97.8, 95.2, and 95.9% identity to the corresponding regions of EV68 prototype strain Fermon. The amino acid identities were 100 and 98.1% for the products of the two capsid regions and 97.9% for 3D RNA polymerase. Antigenic cross-reaction between HRV87 and EV68 was indicated by microneutralization with monotypic antisera. Phylogenetic analysis showed definite clustering of HRV87 and EV68 with EV70 for all sequences examined. Both HRV87 and EV68 were shown to be acid sensitive by two different assays, while EV70 was acid resistant, which is typical of enteroviruses. The cytopathic effect induced by HRV87 or EV68 was inhibited by monoclonal antibodies to the decay-accelerating factor known to be the receptor of EV70. We conclude that HRV87 and EV68 are strains of the same picornavirus serotype presenting features of both rhinoviruses and enteroviruses.
Comparison of Serum Hepatitis C Virus RNA and Core Antigen Concentrations and Determination of Whether Levels Are Associated with Liver Histology or Affected by Specimen Storage Time
Authors: Lagging L. Martin , Garcia Clementina E. , Westin Johan , Wejstål Rune , Norkrans Gunnar , Dhillon Amar P. , Lindh Magnus 
Published: October 2002

ABSTRACT

An enzyme immunoassay has recently been developed for the hepatitis C virus (HCV) core antigen. To evaluate the possible association between core antigen and HCV RNA levels with regards to the change in liver histology over time as well as study the effect of duration of storage on viral load results, sequential sera were analyzed from 45 patients with chronic HCV infection who had undergone two or more liver biopsies. A relatively strong association was found between the core antigen and HCV RNA concentrations (

r
s

= 0.8), with a core antigen level of 1 pg/ml corresponding to approximately 1,000 IU/ml. All 42 sera with detectable HCV RNA at the time of the second biopsy had core antigen concentrations above 1 pg/ml, and the three sera without detectable HCV RNA had concentrations below 1 pg/ml. No association was found between HCV RNA or core antigen levels and the stage of fibrosis in biopsy samples, progression of fibrosis, necro-inflammatory grade, steatosis, genotype, alanine aminotransferase level, or alcohol consumption. A significant association was demonstrated between the storage time of the samples and both the HCV RNA and core antigen concentrations. The median log HCV RNA concentrations (international units/milliliter) were 3.92 for the sera obtained at the time of the first biopsy (median storage time, 13.0 years) and 4.41 for the sera obtained at the time of the second biopsy (median storage time, 6.6 years) compared to 5.96, the median for 102 different routine clinical patient samples.
Identification of the Causative Organism of Tuberculous Lymphadenitis in Ethiopia by PCR
Authors: Kidane Dawit , Olobo Joseph O. , Habte Abebe , Negesse Yohannes , Aseffa Abraham , Abate Getahun , Yassin Mohammed A. , Bereda Kiflu , Harboe Morten 
Published: October 2002

ABSTRACT

Tuberculous lymphadenitis (TBLN) is a common form of extrapulmonary tuberculosis with multiple differential diagnoses. Demonstration of the etiologic agent by smear microscopy or culture of fine needle aspirate (FNA) specimens is often unsuccessful. FNA specimens from 40 patients presenting at a rural health center in South Ethiopia and diagnosed as positive for TBLN on the basis of clinical and cytological criteria were analyzed for mycobacterial DNA by PCR. Thirty (75%) had cervical lymphadenitis and 11 (27.5%) were seropositive for human immunodeficiency virus (HIV). Three primer sets were initially used to identify the causative agent at the genus (antigen 85 complex), complex (IS
6110
insertion sequence), and species (
pncA
gene and allelic variation) levels. Among the forty TBLN cases, 35 (87.5%) were positive by PCR at the genus and complex levels. Based on PCR for detection of allelic variation at position 169, 24 (68.6%) of the 35 were positive for
Mycobacterium tuberculosis
and 6 (17.1%) were positive for
M. bovis
. These six were positive in additional PCR assays using the JB21-JB22 primer set, which is highly specific for
M. bovis.
Five (14.1%) showed amplification for both
M. tuberculosis
and
M. bovis
with the allele-specific primer set
.
Cooccurrence of pyrazinamide (PZA)-sensitive and -resistant
M. tuberculosis
in those five cases was indicated, since all were negative in assays with the JB21-JB22 primer set. This feature was seen in 3 of 11 HIV-positive and 2 of 29 HIV-negative individuals (
P
< 0.001). Conclusion: among 35 PCR-positive cases of TBLN from southern Ethiopia, 29 (82.9%) were caused by
M. tuberculosis
and six (17.1%) were caused by
M. bovis.
Serological Diagnosis of Ovine Enzootic Abortion by Enzyme-Linked Immunosorbent Assay with a Recombinant Protein Fragment of the Polymorphic Outer Membrane Protein POMP90 ofChlamydophila abortus
Authors: Longbottom David , Fairley Susan , Chapman Stephanie , Psarrou Evgenia , Vretou Evangelia , Livingstone Morag 
Published: October 2002

ABSTRACTOvine enzootic abortion (OEA) resulting from infection of sheep and goats withChlamydophila abortusis of major economic importance worldwide. Over the last 50 years the serological diagnosis of infection has been based mainly on the complement fixation test (CFT), which lacks both sensitivity and specificity because of cross-reactive antibodies to other gram-negative bacteria, including another common chlamydial pathogen of sheep,Chlamydophila pecorum. In the present study, a series of overlapping recombinant antigens representing the polymorphic outer membrane protein POMP90 ofC. abortuswas assessed by enzyme-linked immunosorbent assay (ELISA) with a panel of 143 serum samples from sheep experimentally infected withC. abortus, from sheep clinically free of OEA, and from specific-pathogen-free lambs experimentally infected with different subtypes ofC. pecorum. The results were compared to those obtained by CFT and another recently described test, an indirect ELISA (iELISA) with the recombinant OMP91B (rOMP91B) fragment (rOMP91B iELISA) (D. Longbottom, E. Psarrou, M. Livingstone, and E. Vretou, FEMS Microbiol. Lett. 195:157-161, 2001). The rOMP90-3 and rOMP90-4 ELISAs were identified as being more sensitive and specific than CFT. Assays with both fragments were evaluated further with a panel of 294 field serum samples from flocks with documented histories of abortion, from flocks with no clinical histories of abortion but which had a high proportion of samples seropositive by CFT, and from animals with no histories of abortion but from which variousC. pecorumsubtypes had been isolated. ELISAs with both POMP90 fragments outperformed CFT with serum samples fromC. pecorum-infected animals, producing no false-positive results. However, the ELISA with the rOMP90-4 fragment appeared to be more sensitive than the one with rOMP90-3, as it identified more of the OEA-positive samples. The ELISA with the rOMP90-4 fragment was also able to identify apparently healthy animals that were infected with an enteric strain ofC. abortusin flocks that were probably infected with both entericC. abortusandC. pecorumstrains. The identification of animals infected with entericC. abortusis extremely important in controlling the spread of OEA. Overall, the new rOMP90-4 ELISA was found to be a more sensitive and specific test than CFT for differentiating animals infected withC. abortusfrom those infected withC. pecorum.
Development of a Peptide-Mediated Capture PCR for Detection of
Mycobacterium avium
subsp.
paratuberculosis
in Milk
Authors: Stratmann Janin , Strommenger Birgit , Stevenson Karen , Gerlach Gerald-F. 
Published: October 2002

ABSTRACT

Based on phage display technology, a peptide-mediated magnetic separation technique was developed to facilitate selective isolation of
Mycobacterium avium
subsp.
paratuberculosis
(
M. paratuberculosis
) from bulk milk of naturally infected dairy herds. Nine recombinant bacteriophages binding to
M. paratuberculosis
were isolated from a commercial phage-peptide library encoding random 12-mer peptides. Nucleotide sequencing revealed the deduced sequence of the binding peptides. One peptide with the sequence NYVIHDVPRHPA, designated aMP3, was chemically synthesized with an amino-terminal biotin residue attached via an amino-hexacarbonic acid spacer molecule. Paramagnetic beads coated with the phage or with peptide aMP3 enabled the capture of
M. paratuberculosis
from milk. Combining this peptide-mediated magnetic separation with an IS
Mav2
-based PCR allowed the detection of
M. paratuberculosis
in artificially spiked milk down to a concentration of 10
1
ml
−1
. Experiments using milk from naturally infected cows and bulk milk samples from infected herds demonstrated that the peptide-mediated capture PCR is sufficiently sensitive to detect single strong shedders in pooled milk samples. The method, for the first time, applies phage display technology to microbial diagnostics and has potential value as a completely standardizable tool for the routine
M. paratuberculosis
screening of bulk milk samples at acceptable costs.
Improved Monitoring of Cytomegalovirus Infection after Allogeneic Hematopoietic Stem Cell Transplantation by an Ultrasensitive Plasma DNA PCR Assay
Authors: Kaiser Laurent , Perrin Luc , Chapuis Bernard , Hadaya Karine , Kolarova Lenka , Deffernez Christelle , Huguet Saadia , Helg Claudine , Wunderli Werner 
Published: October 2002

ABSTRACTCytomegalovirus (CMV) DNA amplification assays in plasma have shown limited sensitivity compared to the detection of pp65 antigen in leukocytes. Our goal was to increase the sensitivity of a commercial CMV DNA PCR quantitative assay. After modification, the new assay was able to reproducibly detect 20 CMV DNA copies/ml of plasma. We compared this new ultrasensitive PCR assay with the standard PCR and the pp65 test for CMV detection and quantification in 22 consecutive allogeneic hematopoietic stem cell recipients. CMV infection or reactivation was detected in 84 of 319 (26%) samples by the ultrasensitive PCR assay compared to 38 of 319 (12%) samples by the pp65 assay (P< 0.01). All samples positive by the pp65 assay were positive by the ultrasensitive PCR, and CMV episodes were detected on average 4 days earlier and 7 days later than the first and the last pp65-positive test, respectively. In addition, during CMV episodes, the ultrasensitive assay identified positive samples that were inconsistently detected by the pp65 assay. The ultrasensitive assay was also much more sensitive than the standard PCR, with 26 versus 12% of CMV DNA-positive samples (P< 0.01). This assay improved the monitoring of CMV infection or reactivation in hematopoietic allogeneic stem cell recipients.
OspE-Related, OspF-Related, and Elp Lipoproteins Are Immunogenic in Baboons Experimentally Infected withBorrelia burgdorferiand in Human Lyme Disease Patients
Authors: Hefty P. Scott , Brooks Chad S. , Jett Amy M. , White Gary L. , Wikel Stephen K. , Kennedy Ronald C. , Akins Darrin R. 
Published: October 2002

ABSTRACTPresently, the rhesus macaque is the only nonhuman primate animal model utilized for the study of Lyme disease. While this animal model closely mimics human disease, rhesus macaques can harbor the herpes B virus, which is often lethal to humans; macaques also do not express the full complement of immunoglobulin G (IgG) subclasses found in humans. Conversely, baboons contain the full complement of IgG subclasses and do not harbor the herpes B virus. For these reasons, baboons have been increasingly utilized as the basis for models of infectious diseases and studies assessing the safety and immunogenicity of new vaccines. Here we analyzed the capability of baboons to become infected withBorrelia burgdorferi, the agent of Lyme disease. Combined culture and PCR analyses of tick- and syringe-infected animals indicated that baboons are a sufficient host forB. burgdorferi. Analysis of the antibody responses in infected baboons over a 48-week period revealed that antibodies are generated early during infection against many borrelial antigens, including the various OspE, OspF, and Elp paralogs that are encoded on the ubiquitous 32-kb circular plasmids (cp32s). By using the baboon sera generated by experimental infection it was determined that a combination of two cp32-encoded lipoproteins, OspE and ElpB1, resulted in highly specific and sensitive detection ofB. burgdorferiinfection. An expanded analysis, which included 39 different human Lyme disease patients, revealed that a combination of the OspE and ElpB1 lipoproteins could be the basis for a new serodiagnostic assay for Lyme disease. Importantly, this novel serodiagnostic test would be useful independent of prior OspA vaccination status.
Virus Diversity in a Winter Epidemic of Acute Diarrhea in France
Authors: Chikhi-Brachet Roxane , Bon Fabienne , Toubiana Laurent , Pothier Pierre , Nicolas Jean-Claude , Flahault Antoine , Kohli Evelyne 
Published: October 2002

ABSTRACTIn France, an epidemic peak of acute diarrhea is observed each winter. Previous results suggested a viral etiology for these winter epidemics. We investigated the role of enteric viruses in acute diarrhea and their molecular diversity. One hundred sixty-one patients with acute diarrhea and 45 healthy patients (controls) from the general population were given a standardized questionnaire between December 1998 and May 1999. Stool specimens were screened for group A and C rotaviruses, human caliciviruses, astroviruses, and adenovirus types 40 and 41 by reverse transcription-PCR and/or enzyme immunoassay. Virologic analysis was positive for 63 cases (39%). Caliciviruses and group A rotaviruses were the most frequent (19 and 17% of cases, respectively). Two control stool specimens were found positive for group A rotavirus, and one was found positive for astrovirus. Molecular characterization of the strains disclosed a cocirculation of P[8],G1, P[8],G4, and P[4],G2 rotaviruses; type 1, 2, 3, 4, and 8 astroviruses; and Sapporo-like and Norwalk-like human caliciviruses. These four types of viruses accounted for an attributable risk of acute diarrhea of 34.7% for the general population, under the assumption of a causal role of these viruses.
Disseminated Mycotic Infection Caused by
Colletotrichum acutatum
in a Kemp's Ridley Sea Turtle (
Lepidochelys kempi
)
Authors: Manire Charles A. , Rhinehart Howard L. , Sutton Deanna A. , Thompson Elizabeth H. , Rinaldi Michael G. , Buck John D. , Jacobson Elliott 
Published: October 2002

ABSTRACT

Colletotrichum acutatum
is a cosmopolitan plant pathogen with a wide host range. While the organism's phytopathogenic potential has been well documented, it has never been reported as an etiologic agent of disease in either animals or humans. In this case, a juvenile Kemp's ridley sea turtle,
Lepidochelys kempi
, probably with immune compromise following cold stunning (extended hypothermia), developed a disseminated mycotic infection in the lungs and kidneys. Prophylactic treatment with oral itraconazole did not prevent or cure the infection. This report of a
Colletotrichum acutatum
infection in an animal extends the range of disease caused by this organism beyond that of a phytopathogen.
Presumptive Identification ofCandida kefyron Levine Formulation of Eosin Methylene Blue Agar
Authors: Munson Erik L. , Troy Dennis R. , Weber Joanne K. , Messer Shawn A. , Pfaller Michael A. 
Published: October 2002

ABSTRACTThree hundred thirty-one yeast and yeast-like isolates were cultivated on eosin methylene blue agar. While the sensitivity rate forCandida kefyrisolates producing a metallic green sheen was 81.8%, the high positive predictive value (100%) for yeast isolates with this phenotype belonging toC. kefyrsuggests that these isolates can be presumptively identified asC. kefyr.
Modification of Dienes Mutual Inhibition Test for Epidemiological Characterization of
Pseudomonas aeruginosa
Isolates
Authors: Munson Erik L. , Pfaller Michael A. , Doern Gary V. 
Published: October 2002

ABSTRACT

Pseudomonas aeruginosa
is an important cause of community-associated and nosocomial infections related to exposure to aqueous environments. Such infections often occur in the setting of a common-source outbreak, in which case epidemiological characterization of isolates may be necessary. In this preliminary study, a modification of the Dienes mutual inhibition test, ordinarily used to assess the relatedness of swarming
Proteus mirabilis
strains, was used to study 15
P. aeruginosa
isolates, with the results compared to those obtained by ribotype analysis. Complete concordance was noted between the results of the Dienes test and those of ribotyping. These observations suggest that further studies are warranted to assess the utility of the modified Dienes test as a simple, inexpensive, and reliable means for epidemiological typing of
P. aeruginosa
.
Dissemination of New Methicillin-Resistant
Staphylococcus aureus
Clones in the Community
Authors: Okuma Keiko , Iwakawa Kozue , Turnidge John D. , Grubb Warren B. , Bell Jan M. , O'Brien Frances G. , Coombs Geoffrey W. , Pearman John W. , Tenover Fred C. , Kapi Maria , Tiensasitorn Chuntima , Ito Teruyo , Hiramatsu Keiichi 
Published: October 2002

ABSTRACT

Multiple methicillin-resistant
Staphylococcus aureus
(MRSA) clones carrying type IV staphylococcal cassette chromosome
mec
were identified in the community-acquired MRSA strains of both the United States and Australia. They multiplied much faster than health-care-associated MRSA and were resistant to fewer non-beta-lactam antibiotics. They seem to have been derived from more diverse
S. aureus
populations than health-care-associated MRSA strains.
Direct Detection and Identification of African Trypanosomes by Fluorescence In Situ Hybridization with Peptide Nucleic Acid Probes
Authors: Radwanska M. , Magez S. , Perry-O'Keefe H. , Stender H. , Coull J. , Sternberg J. M. , Büscher P. , Hyldig-Nielsen J. J. 
Published: October 2002

ABSTRACT

We have developed a rapid and easy to perform fluorescence in situ hybridization test that allows specific identification of trypanosomes from the subgenus
Trypanozoon
, using peptide nucleic acid probes. Probes were designed to target subgenus-specific sequences on the multiple-copy 18S rRNA, greatly facilitating the detection of a single trypanosome.
Detection of Oxazolidinone-Resistant
Enterococcus faecalis
and
Enterococcus faecium
Strains by Real-Time PCR and PCR-Restriction Fragment Length Polymorphism Analysis
Authors: Woodford Neil , Tysall Luke , Auckland Cressida , Stockdale Mark W. , Lawson Andrew J. , Walker Rachel A. , Livermore David M. 
Published: October 2002

ABSTRACT

A real-time PCR assay identified linezolid-resistant
Enterococcus faecalis
and
Enterococcus faecium
isolates with a G2576U rRNA mutation. PCR-restriction fragment length polymorphism analysis of ribosomal DNA amplicons with
Nhe
I also detected this mutation. Both assays detected isolates heterozygous at this position. Recognition of isolates with what is presently the most frequent oxazolidinone resistance mutation may aid surveillance and individual case management.
Cryopreserved Cell Monolayers for Rapid Detection of Herpes Simplex Virus and Influenza Virus
Authors: Huang Yung T. , Yan Huimin , Sun Yan , Jollick, Joseph A. , Baird Heather 
Published: October 2002

ABSTRACT
Cryopreserved cell monolayers are a new cell culture technology intended to ensure the availability of cells in the laboratory for virus detection. Two cryopreserved cell monolayers, ELVIS for the detection of herpes simplex virus (HSV) and R-Mix for the detection of influenza virus, were evaluated. The results indicated that fresh and cryopreserved cell monolayers are comparable in sensitivity for the detection of HSV and influenza virus. The cells retain the same level of sensitivity for up to 4 months at −80°C.
Real-Time PCR for Detection and Differentiation of Gram-Positive and Gram-Negative Bacteria
Authors: Klaschik Sven , Lehmann Lutz E. , Raadts Ansgar , Book Malte , Hoeft Andreas , Stuber Frank 
Published: October 2002

ABSTRACT
We developed a consensus real-time PCR protocol that enables us to detect spiked bacterial 16S DNA from specimens such as water, urine, plasma, and sputum. The technique allows an exact Gram stain classification of 17 intensive care unit-relevant bacteria by means of fluorescence hybridization probes. All tested bacteria were identified correctly, and none gave a false-positive signal with the opposite Gram probe.
Rapid Identification and Differentiation of
Candida albicans
and
Candida dubliniensis
by Capillary-Based Amplification and Fluorescent Probe Hybridization
Authors: Selvarangan Rangaraj , Limaye Ajit P. , Cookson Brad T. 
Published: October 2002

ABSTRACT

We developed a rapid genotypic assay to differentiate the germ tube-positive yeasts
Candida albicans
and
Candida dubliniensis
. Fluorescently labeled nucleic acid probe binding and subsequent denaturation from the target site in the PCR amplicons produced characteristic peak melting temperatures (

T
m

) that identified each species. Peak

T
m

s of
C. albicans
(
n
= 69) and
C. dubliniensis
(
n
= 28) isolates produced in the presence of their respective probes were 61.04 ± 0.64°C and 60.52 ± 1.01°C (averages ± standard deviations). No signal was generated when the
C. albicans
or
C. dubliniensis
probes were tested against DNA from their counterparts. Both probes reacted with
Candida tropicalis
DNA, but the

T
m

was 51.85 ± 0.05°C with the
C. albicans
probe and 51.92 ± 0.10°C with the
C. dubliniensis
probe, differentiating
C. tropicalis
DNA from
C. albicans
and
C. dubliniensis.
A novel hybrid probe was designed to identify both species in a single reaction based on a 4°C difference in peak

T
m

s. Our assay is rapid (≤2 h) and allows reliable detection and differentiation of the two germ tube-positive
Candida
spp.
Rapid Assessment of Phenotypic Resistance to Protease Inhibitors in Human Immunodeficiency Virus Type 1 Group O
Authors: Rodés Berta , Poveda Eva , Soriano Vincent 
Published: October 2002

ABSTRACT
A bacteriophage lambda-based method was used to investigate the development of resistance to protease inhibitors (PI) in one subject infected with human immunodeficiency virus (HIV) type 1 group O who underwent multiple treatment regimens over a period of 4 years. A reduction in the susceptibility to indinavir of 6-fold and a reduction in the susceptibility to saquinavir of 24-fold were recognized after long exposure to these drugs with respect to baseline. The emergence of PI resistance corresponded to the selection of amino acid changes L10V, G48M, F53L, I54V, and L90M at the protease. The results were concordant with those obtained by a drug susceptibility assay with primary HIV isolates.