bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of a Repetitive Extragenic Palindromic Sequence-Based PCR Method and Clinical and Microbiological Methods for Determining Strain Sources in Cases of Nosocomial
Acinetobacter baumannii
Bacteremia
Authors: Martín-Lozano David , Cisneros José Miguel , Becerril Berta , Cuberos Lucila , Prados Trinidad , Ortíz-Leyba Carlos , Cañas Elías , Pachón Jerónimo 
Published: November 2002

ABSTRACT

Using a repetitive extragenic palindromic PCR (REP-PCR), we genotypically characterized strains causing nosocomial
Acinetobacter baumannii
infections and analyzed the source of bacteremia in 67 patients from an institution in which infections by this bacterium were endemic. Six different genotypes were found, including 21, 27, 3, 9, 3, and 4 strains. The probable source of bacteremia, according to clinical and/or microbiological criteria, was known in 42 patients (63%): respiratory tract (
n
= 19), surgical sites (
n
= 12), intravascular catheters (
n
= 5), burns (
n
= 3), and urinary tract (
n
= 3). The definite source of bacteremia, according to REP-PCR, could be established in 30 (71%) out of the 42 patients with strains from blood and other sites; in these cases clinical and microbiological criteria for the source of bacteremia were thus confirmed. In the remaining 12 patients (29%) the probable source was refuted by the REP-PCR method. The definite sources of bacteremia according to genotype were as follows: respiratory tract in 13 patients (31%), surgical sites in 8 (19%), intravascular catheters in 4 (9%), burns in 3 (7%), and urinary tract in 2 (5%). A comparison of strains from blood cultures and other sites with regard to their REP-PCR and antimicrobial resistance profiles was also made. Taking the REP-PCR as the “gold standard,” the positive predictive value of antibiotype was 77% and the negative predictive value was 42%. In summary, the utility of the diagnosis of the source of nosocomial
A. baumannii
bacteremia using clinical and/or microbiological criteria, including antibiotyping, is limited, as demonstrated by REP-PCR.
Importance of Sample Preparation for Molecular Diagnosis of Lyme Borreliosis from Urine
Authors: Bergmann A. R. , Schmidt B. L. , Derler A.-M. , Aberer E. 
Published: November 2002

ABSTRACT

Urine PCR has been used for the diagnosis of
Borrelia burgdorferi
infection in recent years but has been abandoned because of its low sensitivity and the irreproducibility of the results. Our study aimed to analyze technical details related to sample preparation and detection methods. Crucial for a successful urine PCR were (i) avoidance of the first morning urine sample; (ii) centrifugation at 36,000 ×
g
; and (iii) the extraction method, with only DNAzol of the seven different extraction methods used yielding positive results with patient urine specimens. Furthermore, storage of frozen urine samples at −80°C reduced the sensitivity of a positive urine PCR result obtained with samples from 72 untreated erythema migrans (EM) patients from 85% in the first 3 months to <30% after more than 3 months. Bands were detected at 276 bp on ethidium bromide-stained agarose gels after amplification by a nested PCR. The specificity of bands for 32 of 33 samples was proven by hybridization with a GEN-ETI-K-DEIA kit and for a 10 further positive amplicons by sequencing. By using all of these steps to optimize the urine PCR technique,
B. burgdorferi
infection could be diagnosed by using urine samples from EM patients with a sensitivity (85%) substantially better than that of serological methods (50%). This improved method could be of future importance as an additional laboratory technique for the diagnosis of unclear, unrecognized borrelia infections and diseases possibly related to Lyme borreliosis.
Clinical
Escherichia coli
Strains Carrying
stx
Genes:
stx
Variants and
stx
-Positive Virulence Profiles
Authors: Eklund Marjut , Leino Kirsikka , Siitonen Anja 
Published: November 2002

ABSTRACT

Altogether, 173 Shiga toxin-producing
Escherichia coli
(STEC) serotype O157 (
n
= 111) and non-O157 (
n
= 62) isolates from 170 subjects were screened by PCR-restriction fragment length polymorphism for eight different
stx
genes. The results were compiled according to serotypes, phage types of O157, production of Stx toxin and enterohemolysin, and the presence of
eae
. The
stx
genes occurred in 11 combinations; the most common were
stx
2
with
stx
2c
(42%),
stx
2
alone (21%), and s
tx
1
alone (16%). Of the O157 strains, 64% carried
stx
2
with
stx
2c
versus 2% of the non-O157 strains (
P
< 0.001). In the non-O157 strains, the prevailing gene was s
tx
1
(99% versus 1% in O157 strains;
P
< 0.001). In addition, one strain (O Rough:H4:
stx
2c
) which has not previously been described as associated with hemolytic-uremic syndrome (HUS) was found. Ten
stx
-positive virulence profiles were responsible for 71% of all STEC infections. Of these profiles, five accounted for 71% of the 21 strains isolated from 20 patients with HUS or thrombotic thrombocytopenic purpura (TTP). The strains having the virulence profile that caused mainly HUS or TTP or bloody diarrhea produced Stx with titers of ≥1:128 (90%) more commonly than did other strains (51%;
P
< 0.001). These strains were also more commonly enterohemolytic (98% versus 68% for other strains;
P
< 0.001) and possessed the
eae
gene (100%) more commonly than did other strains (74%;
P
< 0.001). A particular virulence profile, O157:H7:PT2:
stx
2
:
stx
2c
:
eae
:Ehly, was significantly more frequently associated with HUS and bloody diarrhea than were other profiles (
P
= 0.02) and also caused the deaths of two children. In this study, the risk factors for severe symptoms were an age of <5 years and infection by the strain of O157:H7:PT2 mentioned above.
Evaluation of a New Rapid Test for the Combined Detection of Hepatitis B Virus Surface Antigen and Hepatitis B Virus e Antigen
Authors: Clement F. , Dewint P. , Leroux-Roels G. 
Published: November 2002

ABSTRACT

There are about 350 million chronic hepatitis B virus (HBV) carriers worldwide. A proactive approach to the management of this disease is likely to reduce the morbidity and mortality caused by HBV. This study aimed to evaluate the diagnostic performance of a novel tool for discriminating between infected and noninfected subjects, the hepatitis B sAg/eAg test (Binax Inc., Portland, Maine). The test is designed to rapidly and accurately detect both the HBV surface antigen (HBsAg) and the HBV e antigen (HBeAg). A cohort of 942 subjects was tested. The serum clinical sensitivity of the hepatitis B sAg/eAg test was 99.75 and 96.37% for HBsAg and HBeAg, respectively. Serum clinical specificity was 99.32% for HBsAg and 98.99% for HBeAg. Analytical sensitivity was satisfactory for the purposes of population screening. Visual evaluation showed that the test signals were stable for at least 3 h after the recommended evaluation time. No interference or cross-reactivity was observed with known interfering substances and virologic markers. These results indicate that the hepatitis B sAg/eAg test is well suited to the accurate detection of HBV carriers. In addition to the good clinical specificity and sensitivity of this test, its stability and user-friendly design mean that a correct performance, even under field conditions, is highly likely. Consequently, the hepatitis B sAg/eAg test has the potential to identify subjects who require HBV vaccination (HBsAg
−
and HBeAg
−
) and HBV-infected individuals who might benefit most from antiviral therapy (HBsAg
+
and HBeAg
+
).
Use of Subtractive Hybridization To Identify a Diagnostic Probe for a Cystic Fibrosis Epidemic Strain of
Pseudomonas aeruginosa
Authors: Parsons Yasmin N. , Panagea Stavroula , Smart Catherine H. M. , Walshaw Martin J. , Hart C. Anthony , Winstanley Craig 
Published: November 2002

ABSTRACT

A multiresistant strain of
Pseudomonas aeruginosa
is widespread among cystic fibrosis (CF) patients attending clinics in Liverpool, United Kingdom. Suppression subtractive hybridization was used to identify sequences present in the Liverpool CF epidemic strain but absent from strain PAO1. Using dot blot and PCR amplification assays, the prevalence of such sequences among a panel of CF isolates was determined. Several sequences were found only in the Liverpool epidemic strain. Some sequences were present in the Liverpool epidemic strain and in a minority of other isolates, including sequences with homology to genes implicated in O6 serotype and siderophore production. The Liverpool epidemic strain and 81% of nonepidemic isolates contained a sequence identified as part of the PAGI-1 genomic island. Other strains implicated in epidemic spread, which were from Manchester, United Kingdom, and Melbourne, Australia, were also screened. None of the sequences identified was present in the Manchester strain. However, one of two Melbourne strains contained some of the sequences found in the Liverpool epidemic strain. All isolates implicated in epidemic spread and 76% of sporadic isolates contained the
exoS
gene. A sequence present in all isolates of the Liverpool epidemic strain was used to develop a diagnostic PCR test for identification of the strain from colonies or directly from sputum samples.
Bartonella henselaeandBartonella elizabethaeas Potential Canine Pathogens
Authors: Mexas Angela M. , Hancock Susan I. , Breitschwerdt Edward B. 
Published: November 2002

ABSTRACTBartonella henselaeorBartonella elizabethaeDNA from EDTA-anticoagulated blood samples obtained from four dogs was amplified and sequenced. The results showed thatB. elizabethaeshould be added to the list ofBartonellaspecies (i.e.,B. vinsoniisubsp.berkhoffii,B. henselae, andB. clarridgeiae) that are currently recognized as infectious agents in dogs. Furthermore, these results may have potential zoonotic implications, particularly if dogs can serve as a previously unrecognized reservoir forB. henselae. Although the clinical relevance of these observations remains to be determined, it is possible that molecular diagnostic techniques such as PCR may help to implicate a spectrum ofBartonellaspp. as a cause of or a cofactor in chronic canine and human diseases of poorly defined causation.
Analysis of the Pathogenetic Basis for Shedding and Transmission of Ovine Gamma Herpesvirus 2
Authors: Hüssy Daniela , Janett Fredi , Albini Sarah , Stäuber Norbert , Thun Rico , Ackermann Mathias 
Published: November 2002

ABSTRACT

Ovine herpesvirus 2 (OvHV-2), a member of the viral subfamily
Gammaherpesvirinae
, shares numerous similarities with human herpesvirus 8 (HHV-8). Both viruses are apathogenic in their healthy original host, may cause lymphoprolipherative diseases, cannot routinely be propagated in cell culture, and may be sexually transmitted. However, the pathways of sexual transmission of these viruses, as well as the underlying pathogenetic dynamics, are not well understood. Organs from naturally OvHV-2-infected, as well as OvHV-2-free, sheep were quantitatively analyzed for OvHV-2 by the DNA amplification techniques. The dynamics of OvHV-2 multiplication and excretion were monitored after experimental infections and, most importantly, subsequent to vasectomy. The OvHV-2 DNA load in various tissues and internal organs was not merely reflecting the viral DNA load in the bloodstream, which suggested compartmentalization of OvHV-2. Moreover, OvHV-2 DNA was detected at several portals for virus shedding, i.e., the respiratory, alimentary, and urogenital tracts. Transient OvHV-2 excretion was detected in ejaculates of experimentally infected rams. Upon vasectomy, OvHV-2 DNA reappeared in the ejaculatory plasma, but the titers did not decline after reaching a peak. Spiking and fractionation experiments revealed an inhibitory activity, associated with the spermatozoa, which was able to suppress detection of viral DNA but which was no longer present in samples from vasectomized animals. Therefore, epidemiological studies on viruses that may be transmitted by the ejaculatory pathway and for whose tracing nucleic acid amplification methods are used, i.e., OvHV-2, HHV-8, and the human immunodeficiency virus, should include vasectomized males.
Endemic Carbapenem Resistance Associated with OXA-40 Carbapenemase among
Acinetobacter baumannii
Isolates from a Hospital in Northern Spain
Authors: Lopez-Otsoa F. , Gallego L. , Towner K. J. , Tysall L. , Woodford N. , Livermore D. M. 
Published: November 2002

ABSTRACT

Eighty-two carbapenem-resistant isolates of
Acinetobacter baumannii
from a single hospital in Bilbao were typed into two major clusters and several subclusters. Disk synergy tests and PCR indicated the production of a zinc-independent OXA-class carbapenemase. Sequencing identified this enzyme, OXA-40, as a variant of the OXA-24-OXA-25-OXA-26 cluster.
Performance of the Microscopic Observation Drug Susceptibility Assay in Drug Susceptibility Testing for
Mycobacterium tuberculosis
Authors: Park Walter G. , Bishai William R. , Chaisson Richard E. , Dorman Susan E. 
Published: November 2002

ABSTRACT

The drug susceptibility testing performance of a broth-based method with microscopic reading of bacillary growth, the microscopic observation drug susceptibility (MODS) assay, was compared to that of the reference 7H10 agar method of proportion by using 53 isolates of
Mycobacterium tuberculosis
from persons at risk for multidrug-resistant TB. For isoniazid (0.1 μg/ml) and rifampin (2.0 μg/ml), there was 100% agreement between MODS results read at day 11 and the reference method. Levels of agreement for ethambutol tested at 2.5 and 7.5 μg/ml were 70 and 58%, respectively. Levels of agreement for streptomycin tested at 2.0 and 6.0 μg/ml were 77 and 51%, respectively. For isoniazid and rifampin drug susceptibility testing, MODS is as accurate as and more rapid than the reference method.
Rotavirus Particles Can Survive Storage in Ambient Tropical Temperatures for More than 2 Months
Authors: Fischer Thea K. , Steinsland Hans , Valentiner-Branth Palle 
Published: November 2002

ABSTRACT
Typing and in vitro cultivation of rotavirus-positive human stool samples stored unintentionally at ambient tropical temperatures for 2 1/2 months showed that rotavirus is stable and may still be infectious in vitro. This indicates that stool specimen collection for rotavirus studies can be performed in areas and settings where reliable cold storage is not available. The retained infectivity of rotavirus particles underscores the need for safe systems for disposal of feces, in particular in developing countries where rotavirus is a major cause of childhood mortality.
Overestimation of the Hepatitis C Virus RNA Content of Reference Preparations by the AMPLICOR HCV Monitor Test, Version 2.0
Authors: Pisani Giulio , Cristiano Karen , Wirz Maria , Bisso Guillermo M. , Gentili Giuliano 
Published: November 2002

ABSTRACT
An evaluation of the AMPLICOR hepatitis C virus (HCV) monitor test, version 2.0 (Roche Diagnostics), was carried out to investigate whether this test overestimates the HCV RNA content of reference preparations. Satisfactory accuracy was observed when the World Health Organization HCV international standard was included in the assay and a modified formula was used to calculate the viral content.
Are There Naturally Occurring Pleomorphic Bacteria in the Blood of Healthy Humans?
Authors: McLaughlin Richard W. , Vali Hojatollah , Lau Peter C. K. , Palfree Roger G. E. , De Ciccio Angela , Sirois Marc , Ahmad Darakhshan , Villemur Richard , Desrosiers Marcel , Chan Eddie C. S. 
Published: November 2002

ABSTRACT

Dark-field microscopy of blood from healthy individuals revealed the existence of pleomorphic microorganisms. These bacteria exhibited limited growth and susceptibility to antibiotics and could be detected by fluorescent in situ hybridization and flow cytometry. They were further characterized by analysis of their 16S rRNA and
gyrB
genes.
Characterization of a Toxigenic
Vibrio cholerae
O139 Strain Belonging to a New Ribotype and Isolated from a Diarrheal Patient
Authors: Bhanumathi R. , Sabeena F. , Isac Sree Renjini , Radhakutty G. , Singh D. V. 
Published: November 2002

ABSTRACT

We characterized a
Vibrio cholerae
O139 strain isolated from a diarrheal patient admitted to Taluk Hospital, Cherthala, Alleppey, Kerala, India, on 9 June 2000. The
V. cholerae
O139 strain possesses the core of the CTX genetic element, colonization toxin-coregulated pilus, the adherence outer membrane protein, and the central regulatory protein encoded by
toxR
and produces cholera toxin (200 pg/ml). We provide molecular evidence showing that toxigenic
V. cholerae
O139 strain ALO95 belongs to a distinct genotype characterized by a unique ribotype designated B-VII and has a unique enterobacterial repetitive intergenic consensus sequence PCR fingerprint profile designated E-V.
Molecular Epidemiology of Tuberculosis in a Rural Area of High Prevalence in South India: Implications for Disease Control and Prevention
Authors: Narayanan Sujatha , Das Sulochana , Garg Renu , Hari Lalitha , Rao Vijay Bhaskara , Frieden Thomas R. , Narayanan P. R. 
Published: November 2002

ABSTRACT

Molecular and conventional epidemiologic techniques were used to study the mechanisms and risk factors for tuberculosis transmission in a rural area with high prevalence in south India. Restriction fragment length polymorphism analysis with IS
6110
and direct repeat probes was performed with 378
Mycobacterium tuberculosis
isolates from patients. Forty-one percent of
M. tuberculosis
isolates harbored a single copy of IS
6110
. Of 378 patients, 236 had distinct strains; 142 (38%) shared a strain with other patients, indicating recent infection. Older patients, those detected by a house-to-house community survey, and those hospitalized in a sanatorium were more likely to have had a recent infection. These findings suggest that the majority of the tuberculosis cases in south India were due to reactivation; therefore, efforts to control tuberculosis should be sustained.
Effects of fatigue on the torque-velocity relation in muscle
Authors: Spendiff O , Longford N T , Winter E M 
Published: November 2002

Background:The extent to which fatigue is related to the velocity of shortening is still not fully understood.Objectives:To examine the effects of fatigue induced by maximal intensity exercise at different velocities on subsequent torque-velocity relations in muscle.Methods:Ten men (mean (SD) age 25 (3) years; stature 1.78 (0.34) m; body mass 80.6 (14.5) kg) provided written informed consent and, over a five day period in a randomised manner, performed 16 fatiguing bouts of maximal intensity exercise on an isokinetic dynamometer each followed by one repetition maximum action. Concentric peak torque of the preferred leg was assessed at angular velocities of 0.52, 1.05, 2.09, and 3.14 rad/s. Mechanical work performed for all fatiguing bouts was held at 3050–3150 J. After a 45 second rest, a maximum effort was performed at one of the preset velocities. As a baseline, dependent variables of peak torque (N.m), angle of peak torque (rad), and mean torque (N.m) were measured at each velocity. An analysis of covariance was used to compare torque-velocity relations, and one factor, within subject analyses of variance with repeated measures investigated differences from baseline.Results:Torque-velocity relations for all dependent variables did not differ (p>0.05). Similarly, angles at which peak torque occurred were not velocity dependent and did not differ from baseline and subsequent exercise measures (p>0.05). Generally, fatiguing exercise at low velocities led to reductions in peak and mean torque in subsequent exercise at higher velocities (p<0.05).Conclusions:Torque-velocity relations fundamentally remain intact after fatigue induced at any velocity, although the magnitude changes. The results suggest that the greater decline in torque during subsequent exercise at high velocities could be due to greater exhaustion of fatigue sensitive type II fibres, whereas low velocity subsequent exercise is less affected because of the greater use of type I fibres.
Hyperextension trauma to the elbow: radiological and ultrasonographic evaluation in handball goalkeepers
Authors: Popovic N , Lemaire R 
Published: November 2002

Objective:To detect elbow lesions produced by hyperextension in 30 elite handball goalkeepers.Methods:Conventional radiographs, stress radiographs, and ultrasound examination of both elbows were used. As a control group, 30 male volunteers from the general population within the same age group with no history of elbow injury were used.Results:Radiographic findings in the goalkeepers were osteophyte formation in 67%, loose bodies in 5.5%, and periarticular calcification in 5.5%. Significantly greater differences in medial joint space opening between stressed and unstressed elbows were measured in both elbows than in the control group. Ultrasonographic findings showed thickening of the medial collateral ligament in 50%, thickening of the tricipital tendon in 11%, and signs of ulnar neuritis in 22%. An intra-articular effusion was found in 66% and small loose bodies in 33%. No significant differences were found between the dominant and non-dominant elbows at radiological and ultrasound examination. The findings in the control group were normal.Conclusions:The findings support the theory that repetitive hyperextension trauma to the elbow in handball goalkeepers results in pathological changes.
Phase II Study of Oxaliplatin, Fluorouracil, and Folinic Acid in Locally Advanced or Metastatic Gastric Cancer Patients
Authors: Louvet C. , André T. , Tigaud J.M. , Gamelin E. , Douillard J.Y. , Brunet R. , François E. , Jacob J.H. , Levoir D. , Taamma A. , Rougier P. , Cvitkovic E. , de Gramont A. 
Published: November 2002

PURPOSE: To evaluate the efficacy and safety of an oxaliplatin, fluorouracil (5-FU), and folinic acid (FA) combination in patients with metastatic or advanced gastric cancer (M/AGC). PATIENTS AND METHODS: Of the 54 eligible patients with measurable or assessable M/AGC, 53 received oxaliplatin 100 mg/m2 and FA 400 mg/m2 (2-hour intravenous infusion) followed by 5-FU bolus 400 mg/m2 (10-minute infusion) and then 5-FU 3,000 mg/m2 (46-hour continuous infusion) every 14 days. RESULTS: Patients (69% male, 31% female) had a median age of 61 years (range, 31 to 75 years), 89% had a performance status of 0 or 1, 70% had newly diagnosed disease, and 87% had metastatic disease. All had histologically confirmed adenocarcinoma. With a median of three involved organs, disease sites included the lymph nodes (67%), stomach (65%), and liver (61%). A median of 10 cycles per patient and 468 complete cycles were administered. Best responses in the 49 assessable patients were two complete responses and 20 partial responses, giving an overall best response rate of 44.9%. Eight patients underwent complementary treatment with curative intent (six with surgery and two with chemoradiotherapy). Median follow-up, time to progression, and overall survival were 18.6 months, 6.2 months, and 8.6 months, respectively. Grade 3/4 neutropenia, leukopenia, thrombocytopenia, and anemia occurred in 38%, 19%, 4%, and 11% of patients, respectively, and febrile neutropenia occurred in six patients (one episode each). Grade 3 peripheral neuropathy occurred in 21% of patients (oxaliplatin-specific scale). Seven patients withdrew because of treatment-related toxicity. CONCLUSION: This oxaliplatin/5-FU/FA regimen shows good efficacy and an acceptable safety profile in M/AGC patients, and may prove to be a suitable alternative regimen in this indication.