bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of In Situ Methods Used To Detect
Mycobacterium avium
subsp.
paratuberculosis
in Samples from Patients with Crohn's Disease
Authors: Jeyanathan Mangalakumari , Alexander David C. , Turenne Christine Y. , Girard Christiane , Behr Marcel A.
Published: August 2006

In common with other diagnostic tests, detection of mycobacteria in tissue by microscopic examination is susceptible to spectrum bias. Since Crohn's disease is defined by the absence of detectable pathogenic organisms, the use of in situ techniques to search for
Mycobacterium avium
subsp.
paratuberculosis
in Crohn's disease samples requires validation of methods in a paucibacillary setting. To generate paucibacillary infection, C57BL/6 mice were artificially infected with
Mycobacterium avium
subsp.
paratuberculosis
strain K10 and
M. tuberculosis
H37Rv, yielding tissues harboring fewer than one bacillus per oil immersion field. Serial sections of organs were then studied by cell wall-based staining techniques (Ziehl-Neelsen and auramine rhodamine) and nucleic acid-based staining techniques (in situ hybridization [ISH] and indirect in situ PCR [IS PCR]). Microscopic examination and measurement of morphometric parameters of bacilli revealed that for all methods,
Mycobacterium avium
subsp.
paratuberculosis
bacilli were observed to be shorter, smaller, and less rod shaped than
M. tuberculosis
bacilli. Ziehl-Neelsen, auramine rhodamine stains, ISH targeting rRNA, and IS-PCR targeting the IS
900
element afforded comparable sensitivities, but for all methods, visualization of individual bacterial forms required magnification ×1,000. Auramine rhodamine staining and IS-PCR generated positive signals in negative controls, indicating the nonspecificity of these assays. Together, our results indicate that detection of
Mycobacterium avium
subsp.
paratuberculosis
bacilli in tissue requires oil immersion microscopy, that rRNA-ISH provides sensitivity and specificity comparable to those of Ziehl-Neelsen staining, and that the microscopic detection limit for
Mycobacterium avium
subsp.
paratuberculosis
in tissue is governed more by bacterial burden than by staining method.
Invasive Meningococcal Disease in Québec, Canada, Due to an Emerging Clone of ST-269 Serogroup B Meningococci with Serotype Antigen 17 and Serosubtype Antigen P1.19 (B:17:P1.19)
Authors: Law Dennis K. S. , Lorange Manon , Ringuette Louise , Dion Réjean , Giguère Michel , Henderson Averil M. , Stoltz Jan , Zollinger Wendell D. , De Wals Philippe , Tsang Raymond S. W.
Published: August 2006

During periods of endemic meningococcal disease, serogroup B
Neisseria meningitidis
is responsible for a significant percentage of invasive diseases, and no particular clone or strain predominates (F. E. Ashton and D. A. Caugant, Can. J. Microbiol. 47: 293-289, 2001), However, in the winter of 2004 to 2005, a cluster of serogroup B meningococcal disease occurred in one region in the province of Québec, Canada. The
N. meningitidis
strain responsible for this cluster of cases was identified as sequence type ST-269 with the antigenic formula B:17:P1.19. Retrospective analysis of isolates from 2000 onwards showed that this clone first emerged in the province of Québec in 2003. The emergence of this clone of serogroup B meningococci occurred after a mass vaccination against serogroup C
N. meningitidis
, suggesting possible capsule replacement.
Identification of
Acinetobacter
Species and Genotyping of
Acinetobacter baumannii
by Multilocus PCR and Mass Spectrometry
Authors: Ecker Joseph A. , Massire Christian , Hall Thomas A. , Ranken Raymond , Pennella Thuy-Trang D. , Agasino Ivy Cristina , Blyn Lawrence B. , Hofstadler Steven A. , Endy Timothy P. , Scott Paul T. , Lindler Luther , Hamilton Tacita , Gaddy Charla , Snow Kerry , Pe Marie , Fishbain Joel , Craft David , Deye Gregory , Riddell Scott , Milstrey Eric , Petruccelli Bruno , Brisse Sylvain , Harpin Vanessa , Schink Amy , Ecker David J. , Sampath Rangarajan , Eshoo Mark W.
Published: August 2006

Members of the genus
Acinetobacter
are ubiquitous in soil and water and are an important cause of nosocomial infections. A rapid method is needed to genotype
Acinetobacter
isolates to determine epidemiology and clonality during infectious outbreaks. Multilocus PCR followed by electrospray ionization mass spectrometry (PCR/ESI-MS) is a method that uses the amplicon base compositions to genotype bacterial species. In order to identify regions of the
Acinetobacter
genome useful for this method, we sequenced regions of six housekeeping genes (
trpE
,
adk
,
efp
,
mutY
,
fumC
, and
ppa
) from 267 isolates of
Acinetobacter
. Isolates were collected from infected and colonized soldiers and civilians involved in an outbreak in the military health care system associated with the conflict in Iraq, from previously characterized outbreaks in European hospitals, and from culture collections. Most of the isolates from the Iraqi conflict were
Acinetobacter baumannii
(189 of 216 isolates). Among these, 111 isolates had genotypes identical or very similar to those associated with well-characterized
A. baumannii
isolates from European hospitals. Twenty-seven isolates from the conflict were found to have genotypes representing different
Acinetobacter
species, including 8 representatives of
Acinetobacter
genomospecies 13TU and 13 representatives of
Acinetobacter
genomospecies 3. Analysis by the PCR/ESI-MS method using nine primer pairs targeting the most information-rich regions of the
trpE
,
adk
,
mutY
,
fumC
, and
ppa
genes distinguished 47 of the 48
A. baumannii
genotypes identified by sequencing and identified at the species level at least 18
Acinetobacter
species. Results obtained with our genotyping method were essentially in agreement with those obtained by pulse-field gel electrophoresis analysis. The PCR/ESI-MS genotyping method required 4 h of analysis time to first answer with additional samples subsequently analyzed every 10 min. This rapid analysis allows tracking of transmission for the implementation of appropriate infection control measures on a time scale previously not achievable.
Catheter-Related Fungemia Due toCandida thermophila
Authors: Bar-Meir Maskit , Sutton Deanna A. , Wickes Brian , Kurtzman Cletus P. , Goldman Stewart , Zheng Xiaotian
Published: August 2006

We report a case of bloodstream infection caused byCandida thermophila, a yeast not previously associated with human disease. The infection occurred in a 13-year-old boy with medulloblastoma who presented with 1 day of fever. Multiple blood cultures were positive for yeast. Removal of the catheter resulted in prompt resolution of the fever and sterilization of the blood cultures. The species was identified by sequencing domains 1 and 2 of the large subunit rRNA gene. Antifungal susceptibility testing was also performed.
Comparison of Dalbavancin MIC Values Determined by Etest (AB BIODISK) and Reference Dilution Methods Using Gram-Positive Organisms
Authors: Fritsche Thomas R. , Rennie Robert P. , Goldstein Beth P. , Jones Ronald N.
Published: August 2006

While standardized microdilution testing methodologies and quality control ranges exist for the novel glycolipopeptide dalbavancin, no testing methods have been described that are immediately available for routine use in clinical laboratories. In this study, we found that the dalbavancin Etest (AB BIODISK, Solna, Sweden) procedure demonstrated a high degree of agreement (100% within ±2 log
2
dilution steps) with the standardized broth microdilution method, validating the use of the Etest as an alternative test for investigational or clinical purposes following regulatory approval.
Rapid Detection of the
Mycobacterium tuberculosis
Beijing Genotype and Its Ancient and Modern Sublineages by IS
6110
-Based Inverse PCR
Authors: Mokrousov Igor , Jiao Wei Wei , Valcheva Violeta , Vyazovaya Anna , Otten Tatiana , Ly Ho Minh , Lan Nguyen Ngoc , Limeschenko Elena , Markova Nadya , Vyshnevskiy Boris , Shen A Dong , Narvskaya Olga
Published: August 2006

The
Mycobacterium tuberculosis
Beijing genotype strains appear to be hypervirulent and associated with multidrug-resistant tuberculosis. Therefore, the development of a both rapid and simple method to detect the
M. tuberculosis
Beijing genotype is of clinical interest per se. Previously, we described a simple and fast approach to detect the Beijing genotype based on IS
6110
inverse-PCR typing. Here, we evaluated this method against a large, diverse, and recent collection of strains. The study sample included 866
M. tuberculosis
strains representing but not limited to the regions in Russia, Europe, and East Asia where the Beijing genotype is endemic. Based on a spoligotyping method, 408 strains were identified as Beijing genotypes; they were additionally subdivided into ancient and modern sublineages based on the analysis of the NTF locus. All strains were further subjected to the IS
6110
-based inverse PCR. All of the Beijing genotype strains were found to have identical two-band (ancient sublineage) or three-band (modern sublineage) profiles that were easily recognizable and distinct from the profiles of the non-Beijing strains. Therefore, we suggest using IS
6110
-based inverse-PCR typing for the correct identification of the Beijing genotype and its major sublineages. The method is fast and inexpensive and does not require additional experiments but instead is implemented in the routine typing method of
M. tuberculosis
.
Multilocus Sequence Typing of Historical
Burkholderia pseudomallei
Isolates Collected in Southeast Asia from 1964 to 1967 Provides Insight into the Epidemiology of Melioidosis
Authors: McCombie Roberta L. , Finkelstein Richard A. , Woods Donald E.
Published: August 2006

A collection of 207 historically relevant
Burkholderia pseudomallei
isolates was analyzed by multilocus sequence typing (MLST). The strain collection contains environmental isolates obtained from a geographical distribution survey of
B. pseudomallei
isolates in Thailand (1964 to 1967), as well as stock cultures and colony variants from the U.S. Army Medical Research Unit (Malaysia), the Walter Reed Army Institute for Research, and the Pasteur Institute (Vietnam). The 207 isolates of the collection were resolved into 80 sequence types (STs); 56 of these were novel. eBURST diagrams predict that the historical-collection STs segregate into three complexes when analyzed separately. When added to the 760 isolates and 365 STs of the
B. pseudomallei
MLST database, the historical-collection STs cluster significantly within the main complex of the eBURST diagram in an ancestral pattern and alter the
B. pseudomallei
“population snapshot.” Differences in colony morphology among reference isolates were found not to affect the STs assigned, which were consistent with the original isolates. Australian ST84 is likely characteristic of
B. pseudomallei
isolates of Southeast Asia rather than Australia, since multiple environmental isolates from Thailand and Malaysia share this ST with the single Australian clinical isolate in the MLST database. Phylogenetic evidence is also provided suggesting that Australian isolates may not be distinct from those of Thailand, since ST60 is common to environmental isolates from both countries. MLST and eBURST are useful tools for the study of population biology and epidemiology, since they provide methods to elucidate new genetic relationships among bacterial isolates.
Impact of Laboratory Cross-Contamination on Molecular Epidemiology Studies of Tuberculosis
Authors: Martínez Miguel , de Viedma Darío García , Alonso María , Andrés Sandra , Bouza Emilio , Cabezas Teresa , Cabeza Isabel , Reyes Armando , Sánchez-Yebra Waldo , Rodríguez Manuel , Sánchez M. Isabel , Rogado M. Cruz , Fernández Rosa , Peñafiel Teresa , Martínez Juan , Barroso Pilar , Lucerna M. Ángeles , Diez L. Felipe , Gutiérrez Carmelo
Published: August 2006

Laboratory cross-contamination by
Mycobacterium tuberculosis
is known to be responsible for the misdiagnosis of tuberculosis, but its impact on other contexts has not been analyzed. We present the findings of a molecular epidemiology analysis in which the recent transmission events identified by a genotyping reference center were overestimated as a result of unnoticed laboratory cross-contamination in the original diagnostic laboratories.
Epidemiology of Candidemia in Brazil: a Nationwide Sentinel Surveillance of Candidemia in Eleven Medical Centers
Authors: Colombo Arnaldo L. , Nucci Marcio , Park Benjamin J. , Nouér Simone A. , Arthington-Skaggs Beth , da Matta Daniel A. , Warnock David , Morgan Juliette
Published: August 2006

Candidemia studies have documented geographic differences in rates and epidemiology, underscoring the need for surveillance to monitor trends. We conducted prospective candidemia surveillance in Brazil to assess the incidence, species distribution, frequency of antifungal resistance, and risk factors for fluconazole-resistant
Candida
species. Prospective laboratory-based surveillance was conducted from March 2003 to December 2004 in 11 medical centers located in 9 major Brazilian cities. A case of candidemia was defined as the isolation of
Candida
spp. from a blood culture. Incidence rates were calculated per 1,000 admissions and 1,000 patient-days. Antifungal susceptibility tests were performed by using the broth microdilution assay, according to the Clinical and Laboratory Standards Institute guidelines. We detected 712 cases, for an overall incidence of 2.49 cases per 1,000 admissions and 0.37 cases per 1,000 patient-days. The 30-day crude mortality was 54%.
C. albicans
was the most common species (40.9%), followed by
C. tropicalis
(20.9%) and
C. parapsilosis
(20.5%). Overall, decreased susceptibility to fluconazole occurred in 33 (5%) of incident isolates, 6 (1%) of which were resistant. There was a linear correlation between fluconazole and voriconazole MICs (
r
= 0.54 and
P
< 0.001 [Spearman's rho]). This is the largest multicenter candidemia study conducted in Latin America and shows the substantial morbidity and mortality of candidemia in Brazil. Antifungal resistance was rare, but correlation between fluconazole and voriconazole MICs suggests cross-resistance may occur.
Serotype 1-Specific Monoclonal Antibody-Based Antigen Capture Immunoassay for Detection of Circulating Nonstructural Protein NS1: Implications for Early Diagnosis and Serotyping of Dengue Virus Infections
Authors: Xu Hua , Di Biao , Pan Yu-xian , Qiu Li-wen , Wang Ya-di , Hao Wei , He Li-juan , Yuen Kwok-yung , Che Xiao-yan
Published: August 2006

Rapid diagnosis and serotyping of dengue virus (DV) infections are important for timely clinical management and epidemiological control in areas where multiple flaviviruses are endemic. However, the speed and accuracy of diagnosis must be balanced against test cost and availability, especially in developing countries. We developed a specific antigen capture enzyme-linked immunosorbent assay (ELISA) for early detection and serotyping of DV serotype 1 (DV1) by using well-characterized monoclonal antibodies (MAbs) specific to nonstructural protein 1 (NS1) of DV1. With this assay, a total of 462 serum specimens from clinically probable DV1-infected patients during the DV1 epidemic in Guangdong, China, in 2002 and 2003 were analyzed. DV1 NS1 was detectable in blood circulation from the first day up to day 18 after onset of symptoms, with a peak at days 6 to 10. The sensitivity of DV1 NS1 detection in serum specimens with reference to results from reverse transcriptase PCR was 82%, and the specificity was 98.9% with reference to 469 healthy blood donors. No cross-reactions with any of the other three DV serotypes or other closely related members of the genus
Flavivirus
(
Japanese encephalitis virus
and
Yellow fever virus
) were observed when tested with the clinical specimens or virus cultures. These findings suggest that the serotype-specific MAb-based NS1 antigen capture ELISA may be a valuable tool for early diagnosis and serotyping of DV infections, while also providing a standardized assay for the analysis of a great number of clinical samples with convenience and cost-effectiveness.
Molecular Analysis of Fungal Microbiota in Samples from Healthy Human Skin and Psoriatic Lesions
Authors: Paulino Luciana C. , Tseng Chi-Hong , Strober Bruce E. , Blaser Martin J.
Published: August 2006

Psoriasis, a common cutaneous disease of unknown etiology, may be triggered by infections, including those due to fungi. Since the fungal community of human skin is poorly characterized, we aimed to analyze the mycological microbiota in healthy skin and psoriatic lesions. Twenty-five skin samples from five healthy subjects (flexor forearm) and three patients with psoriasis were analyzed using broad-range 18S ribosomal DNA (rDNA) and 5.8S rDNA/internal transcribed spacer 2 (ITS2)Malassezia-specific PCR primers. Broad-range PCR analysis indicated that most organisms resembledMalassezia. Malassezia-specific 5.8S/ITS2 analysis of 1,374 clones identified five species and four unknown phylotypes, potentially representing new species. The species distribution appears largely host specific and conserved in different sites of healthy skin. In three subjects, theMalasseziamicrobiota composition appeared relatively stable over time. Samples ofMalasseziamicrobiota from healthy skin and psoriatic lesions were similar in one patient but substantially different in two others. These data indicate the predominance ofMalasseziaorganisms in healthy human skin, host-specific variation, stability over time, and as yet, no consistent patterns differentiating psoriatic skin from healthy skin.
Evaluation of Two New Immunochromatographic Assays (Rapid U Legionella Antigen Test and SD Bioline Legionella Antigen Test) for Detection of
Legionella pneumophila
Serogroup 1 Antigen in Urine
Authors: Diederen Bram M. W. , Peeters Marcel F.
Published: August 2006

We evaluated two new immunochromatographic assays for their abilities to detect
Legionella pneumophila
serogroup 1 antigen in urine. The results were compared with those obtained by the Binax NOW urinary antigen test. The sensitivities and specificities were estimated to be 71.2% and 96.6%, respectively, for the Rapid U test; 31.5% and 98.9%, respectively, for the SD Bioline test; and 91.8% and 100%, respectively, for the Binax NOW test.
Multiple Cases of Familial Transmission of Community-Acquired Methicillin-Resistant
Staphylococcus aureus
Authors: Huijsdens X. W. , van Santen-Verheuvel M. G. , Spalburg E. , Heck M. E. O. C. , Pluister G. N. , Eijkelkamp B. A. , de Neeling A. J. , Wannet W. J. B.
Published: August 2006

The worldwide emergence of community-acquired methicillin-resistant
Staphylococcus aureus
(CA-MRSA) can have severe public health implications. Familial transmissions of CA-MRSA in The Netherlands were investigated. Among the families studied, two clusters of CA-MRSA could be identified. This report demonstrates that family members can serve as reservoirs of CA-MRSA which may become a serious problem in containing the spread of MRSA.
Clinical Evaluation of a Type III Secretion System Real-Time PCR Assay for Diagnosing Melioidosis
Authors: Meumann Ella M. , Novak Ryan T. , Gal Daniel , Kaestli Mirjam E. , Mayo Mark , Hanson Joshua P. , Spencer Emma , Glass Mindy B. , Gee Jay E. , Wilkins Patricia P. , Currie Bart J.
Published: August 2006

A
Burkholderia pseudomallei
type III secretion system real-time PCR assay was evaluated on clinical specimens in a region where melioidosis is endemic. The PCR was positive in 30/33 (91%) patients with culture-confirmed melioidosis. All six patients with melioidosis septic shock were blood PCR positive, suggesting potential for rapid diagnosis and commencement of appropriate therapy.
Ciprofloxacin Treatment of Bacterial Peritonitis Associated with Chronic Ambulatory Peritoneal Dialysis Caused by
Neisseria cinerea
Authors: Taegtmeyer M. , Saxena R. , Corkill J. E. , Anijeet H. , Parry C. M.
Published: August 2006

Bacterial peritonitis is a well-recognized complication of chronic ambulatory peritoneal dialysis (CAPD) in patients with end-stage renal failure. We present a case of peritonitis due to an unusual pathogen,
Neisseria cinerea
, unresponsive to the standard intraperitoneal (i.p.) vancomycin and gentamicin, which responded rapidly to oral ciprofloxacin.
Identification of Genotypes of
Enterocytozoon bieneusi
from Stool Samples from Human Immunodeficiency Virus-Infected Patients in Thailand
Authors: Leelayoova Saovanee , Subrungruang Ittisak , Suputtamongkol Yupin , Worapong Jeerapun , Petmitr Porntip Chavalitshewinkoon , Mungthin Mathirut
Published: August 2006

We identified genotypes of
Enterocytozoon bieneusi
from 33 stool samples of Thai human immunodeficiency virus (HIV)-infected adult patients. Genotype D was identified at the highest frequency (36.4%), while genotype E was the second most common (15.1%). Genotypes O and
Pig
EBITS 7, previously found only in pigs, were observed in Thai HIV-infected patients. Phylogenetic analysis supported a zoonotic nature for
E. bieneusi
.
Measles Virus-Specific Antibody Levels in Individuals in Argentina Who Received a One-Dose Vaccine
Authors: Argüelles Marcelo H. , Orellana Mariana L. , Castello Alejandro A. , Villegas Guillermo A. , Masini Matilde , Belizan Alejandra L. , González Ayala Silvia , Vera Osmar D. , Glikmann Graciela
Published: August 2006

In spite of active measles virus (MV) vaccination strategies, reemergence continues to occur, impairing global eradication programs. The immune status against measles was evaluated in 350 vaccinated healthy Argentine children and teenagers who received a single dose of the MV Schwarz strain Lirugen vaccine (Aventis Pasteur). Sera were assessed for immunoglobulin G (IgG) antibodies by a commercial enzyme immunoassay (EIA) (Enzygnost; Behring), an in-house EIA, and neutralization EIA. Results obtained with these methods showed a marked decline in IgG level with increasing age. At 1 to 4 years of age, 84% of children had IgG antibodies above 200 mIU/ml, conventionally accepted as protective levels, whereas only 32% of older children and teenagers had antibody levels exceeding 200 mIU/ml. Moreover, the MV IgG content in the teenage group was significantly lower than the IgG antibody level of the group of younger children (
P
< 0.0001). In contrast, screening for IgG antibody levels to inactivated tetanus vaccine showed that, on average, 80% of this population was fully protected and that this high level of protection remained through the teenage years. This study suggests that within this population a considerable proportion of individuals had low measles antibody levels that may be insufficient to protect against reinfections or clinical disease.
Identification of
Acinetobacter baumannii
by Detection of the
bla
OXA-51-like
Carbapenemase Gene Intrinsic to This Species
Authors: Turton Jane F. , Woodford Neil , Glover Judith , Yarde Susannah , Kaufmann Mary E. , Pitt Tyrone L.
Published: August 2006

bla
OXA-51-like
was sought in clinical isolates of
Acinetobacter
species in a multiplex PCR, which also detects
bla
OXA-23-like
and class 1 integrase genes. All isolates that gave a band for
bla
OXA-51-like
identified as
A. baumannii
. This gene was detected in each of 141 isolates of
A. baumannii
but not in those of 22 other
Acinetobacter
species.
Widely Distributed and Predominant CTX-M Extended-Spectrum β-Lactamases in Amsterdam, The Netherlands
Authors: al Naiemi N. , Bart A. , de Jong M. D. , Vandenbroucke-Grauls C. M. , Rietra P. J. G. M. , Debets-Ossenkopp Y. J. , Wever P. C. , Spanjaard L. , Bos A. J. , Duim B.
Published: August 2006

Three hundred sixty
Enterobacteriaceae
and nonfermenting gram-negative bacilli, isolated during one week in May 2004 at five hospitals in Amsterdam, The Netherlands, were evaluated for the presence of extended-spectrum beta-lactamases (ESBLs). A prevalence of 7.8% was found, in contrast to the 1% observed in 1997. CTX-M ESBLs dominated, and four types were identified in 18 isolates.
Multitypic Hepatitis C Virus Infection Identified by Real-Time Nucleotide Sequencing of Minority Genotypes
Authors: Buckton Andrew J. , Ngui Siew-Lin , Arnold Catherine , Boast Katie , Kovacs Joanne , Klapper Paul E. , Patel Bharat , Ibrahim Imad , Rangarajan Savita , Ramsay Mary E. , Teo Chong-Gee
Published: August 2006

The prevalence of concurrent multitypic hepatitis C virus (HCV) infection is uncertain. A sensitive and specific approach to identifying minority HCV genotypes in blood is presented. Following serum extraction and reverse transcription PCR to amplify cDNA originating from the viral 5′ noncoding region, the amplified product mixture was treated with genotype-specific restriction endonuclease to digest the dominant genotype. Residual amplicons were subjected to PCR cloning and then to real-time DNA sequencing using a Pyrosequencer to identify the remaining genotypes. Dilution experiments showed that minority genotypes may be detected when they represent 1:10,000 of the total population and in serum specimens with viral loads as low as 1,000 IU/ml. Of 37 patients with bleeding disorders and 44 injecting drug users, infection by more than one HCV genotype was found in 7 (19%) and 4 (9%) patients, respectively. The low rate of detection in people at high risk of repeated HCV infection suggests that multitypic HCV carriage is uncommon.
Clonality and Antimicrobial Resistance Gene Profiles of Multidrug- Resistant
Salmonella enterica
Serovar Infantis Isolates from Four Public Hospitals in Rio de Janeiro, Brazil
Authors: Fonseca E. L. , Mykytczuk O. L. , Asensi M. D. , Reis E. M. F. , Ferraz L. R. , Paula F. L. , Ng L. K. , Rodrigues D. P.
Published: August 2006

In Brazil,
Salmonella enterica
serovar Infantis resistant to various antimicrobials, including cephalosporins, has been identified as an etiological agent of severe gastroenteritis in hospitalized children since 1994. In this study, 35 serovar Infantis strains, isolated from children admitted to four different Rio de Janeiro, Brazil, hospitals between 1996 and 2001, were characterized by pulsed-field gel electrophoresis (PFGE) and antimicrobial susceptibility testing in order to determine their genetic relatedness and antimicrobial resistance profiles. Thirty-four serovar Infantis strains were resistant to at least two antibiotic classes, and all 35 strains were susceptible to fluoroquinolones, cephamycin, and carbapenem. Extended-spectrum beta-lactamase (ESBL) screening by double-disk diffusion indicated that 32 serovar Infantis strains (91.4%) produced beta-lactamases that were inhibited by clavulanic acid. Antimicrobial resistance gene profiles were determined by PCR for a subset of 11 multidrug-resistant serovar Infantis strains, and putative ESBLs were detected by isoelectric focusing. Ten serovar Infantis strains carried
bla
TEM
,
catI
,
ant(3
"
)Ia
and/or
ant(3
"
)Ib
,
sulI
and/or
sulII
, and
tet
(D) genes as well as an integron-associated
aac(6
′
)-Iq
cassette. Eight strains possessed at least four different beta-lactamases with pI profiles that confirmed the presence of both ESBLs and non-ESBLs. Our PFGE profiles indicated that 33 serovar Infantis strains isolated from Rio de Janeiro hospitals came from the same genetic lineage.
False-Positive Results of Enzyme Immunoassays for Human Immunodeficiency Virus in Patients with Uncomplicated Malaria
Authors: Gasasira Anne F. , Dorsey Grant , Kamya Moses R. , Havlir Diane , Kiggundu Moses , Rosenthal Philip J. , Charlebois Edwin D.
Published: August 2006

Malaria may impact upon human immunodeficiency virus (HIV) test results. We evaluated two HIV enzyme immunoassays (EIAs) by testing 1,965 Ugandans with malaria. We found poor positive predictive values (53% and 76%), particularly with younger age. Combining EIAs eliminated false positives but missed 21% of true positives. Performance of HIV EIAs in malaria may be unsatisfactory.
Concurrent Analysis of Nose and Groin Swab Specimens by the IDI-MRSA PCR Assay Is Comparable to Analysis by Individual-Specimen PCR and Routine Culture Assays for Detection of Colonization by Methicillin-Resistant
Staphylococcus aureus
Authors: Bishop Emma J. , Grabsch Elizabeth A. , Ballard Susan A. , Mayall Barrie , Xie Shirley , Martin Rhea , Grayson M. Lindsay
Published: August 2006

The IDI-MRSA assay (Infectio Diagnostic, Inc., Sainte-Foy, Quebec, Canada) with the Smart Cycler II rapid DNA amplification system (Cepheid, Sunnyvale, CA) appears to be sensitive and specific for the rapid detection of nasal colonization by methicillin-resistant
Staphylococcus aureus
(MRSA). We assessed the sensitivity and specificity of this assay under conditions in which both the nose and cutaneous groin specimens were analyzed together and compared the accuracy of this PCR approach to that when these specimens were tested separately and by culture assays in an inpatient population with known high rates (12 to 15%) of MRSA colonization. Of 211 patients screened, 192 had results assessable by all three methods (agar-broth culture, separate nose and groin IDI-MRSA assay, and combined nose-groin IDI-MRSA assay), with MRSA carriage noted in 31/192 (16.1%), 41/192 (21.4%), and 36/192 (18.8%) patients by each method, respectively. Compared to agar culture results, the sensitivity and specificity of the combined nose-groin IDI-MRSA assay were 88.0% and 91.6%, respectively, whereas when each specimen was processed separately, the sensitivities were 90.0% (nose) and 83.3% (groin) and the specificities were 91.7% (nose) and 90.2% (groin). IDI-MRSA assay of a combined nose-groin specimen appears to have an accuracy similar to that of the current recommended PCR protocol, providing results in a clinically useful time frame, and may represent a more cost-effective approach to using this assay for screening for MRSA colonization.
Molecular Epidemiology of
Neisseria gonorrhoeae
in Amsterdam, The Netherlands, Shows Distinct Heterosexual and Homosexual Networks
Authors: Kolader Marion-Eliëtte , Dukers Nicole H. T. M. , van der Bij Akke K. , Dierdorp Mirjam , Fennema Johan S. A. , Coutinho Roel A. , Bruisten Sylvia M.
Published: August 2006

Molecular typing, added to epidemiological data, can better identify transmission patterns of gonorrhea in Western countries, where the incidence has recently been rising. From September 2002 to September 2003, patients with a laboratory-confirmed diagnosis of gonorrhea at the Clinic for Sexually Transmitted Infections in Amsterdam, The Netherlands, were subjected to a questionnaire pertaining to sexual risk behavior and sexual partners in the 6 months prior to the diagnosis. The
Neisseria gonorrhoeae
isolates were all genotyped using PCR-restriction fragment length polymorphism of the porin and opacity genes. All patients with a completed questionnaire and genotyped isolates were included in the study. We obtained 885
N. gonorrhoeae
isolates from 696 patients that revealed 88 clusters and 46 unique genotypes. Patients infected at multiple anatomical sites with one or more strains and patients infected several times during the study period were shown to pursue high-risk sexual behavior and were considered core groups. There were 11 clusters of ≥20 patients; in seven clusters, 81% to 100% of patients were men who have sex with men (MSM), three clusters contained 87 to 100% heterosexual men and women, and one cluster was formed by equal proportions of MSM and heterosexual male and female patients. However, the various clusters differed in characteristics such as types of coinfections, numbers of sexual partners, Internet use to seek sexual partners, and locations of sexual encounters. Molecular epidemiology of gonococcal isolates in Amsterdam revealed core groups and clusters of MSM and heterosexual patients that probably indicate distinct transmission networks.
Sensitivity Analysis and Potential Uses of a Novel Gamma Interferon Release Assay for Diagnosis of Tuberculosis
Authors: Tsiouris Simon J. , Coetzee David , Toro Patricia L. , Austin Judy , Stein Zena , El-Sadr Wafaa
Published: August 2006

Sputum smears for acid-fast bacilli (AFB) are the primary methods for diagnosis of tuberculosis (TB) in many countries. The tuberculin skin test (TST) is the primary method for diagnosis of latent TB infection (LTBI) worldwide. The poor sensitivity of the former and the poor specificity of the latter warrant the development of new tests and strategies to enhance diagnostic capabilities. We evaluated the sensitivity of an “in-tube” gamma interferon release assay (IGRA) using TB-specific antigens in comparison to the TST and the sputum smear for AFB in TB cases in South Africa. The sensitivity of the IGRA for TB was considered a surrogate of sensitivity in LTBI. Among 154 patients with a positive culture for
Mycobacterium tuberculosis
, the sensitivity of the IGRA for the diagnosis of TB varied by clinical subgroup from 64% to 82%, that of the TST varied from 85% to 94%, and that of two sputum smears for AFB varied from 35% to 53%. The sensitivity of the IGRA in human immunodeficiency virus (HIV)-infected TB cases was 81%. HIV-infected TB patients were significantly more likely to have indeterminate IGRA results and produced quantitatively less gamma interferon in response to TB-specific antigens than HIV-negative TB patients. The overall sensitivity of the TST in all TB cases was higher than that of the IGRA (90% versus 76%, respectively). The combined sensitivities of the TST plus IGRA and TST plus a single sputum smear were 96% and 93%, respectively. The TST combined with IGRA or with a single sputum smear may have a role in excluding the diagnosis of TB in some settings.
Comparison of Real-Time PCR Assays for Monitoring Serum Hepatitis B Virus DNA Levels during Antiviral Therapy
Authors: Hui Chee-Kin , Bowden Scott , Zhang Hai-Ying , Wong Anita , Lewin Sharon , Rousseau Frank , Mommeja-Marin Herve , Lee Nikki P. , Luk John M. , Locarnini Stephen , Leung Nancy , Naoumov Nikolai V. , Lau George K. K.
Published: August 2006

The performance characteristics of the RealART and Molecular Beacons assays were compared with those of the Digene Hybrid Capture II assay (ultrasensitive). The results of the RealART and Digene Hybrid assays were related (
r
= 0.94;
P
< 0.001) and diverged by 2 orders of magnitude. The RealART assay can be used to effectively monitor serum hepatitis B virus DNA levels.
Clonal Emergence of Extended-Spectrum β-Lactamase (CTX-M-2)-Producing
Salmonella enterica
Serovar Virchow Isolates with Reduced Susceptibilities to Ciprofloxacin among Poultry and Humans in Belgium and France (2000 to 2003)
Authors: Bertrand Sophie , Weill François-Xavier , Cloeckaert Axel , Vrints Martine , Mairiaux Eric , Praud Karine , Dierick Katlijne , Wildemauve Christa , Godard Claudine , Butaye Patrick , Imberechts Hein , Grimont Patrick A. D. , Collard Jean-Marc
Published: August 2006

Antibiotic treatment is not required in cases of
Salmonella enterica
gastroenteritis but is essential in cases of enteric fever or invasive salmonellosis or in immunocompromised patients. Although fluoroquinolones and extended-spectrum cephalosporins are the drugs of choice to treat invasive
Salmonella
, resistance to these antibiotics is increasing worldwide. During the period 2000 to 2003, 90
Salmonella enterica
serovar Virchow poultry and poultry product isolates and 11 serovar Virchow human isolates were found to produce an extended-spectrum β-lactamase, CTX-M-2, concomitantly with a TEM-1 β-lactamase. The
bla
CTX-M-2
gene was located on a large conjugative plasmid (>100 kb). Pulsed-field gel electrophoresis indicated a clonal relationship between the poultry and human isolates. All these isolates displayed additional resistance to trimethoprim-sulfamethoxazole and tetracycline as well as a reduced susceptibility to ciprofloxacin (MICs of between 0.5 and 1 μg/ml). CTX-M-2-producing
Salmonella
with a reduced susceptibility to fluoroquinolones constitutes a major concern, since such strains could disseminate on a large scale and jeopardize classical antibiotic therapy in immunocompromised patients.
Effect of Sequence Variation in
Plasmodium falciparum
Histidine- Rich Protein 2 on Binding of Specific Monoclonal Antibodies: Implications for Rapid Diagnostic Tests for Malaria
Authors: Lee Nelson , Baker Joanne , Andrews Kathy T. , Gatton Michelle L. , Bell David , Cheng Qin , McCarthy James
Published: August 2006

The ability to accurately diagnose malaria infections, particularly in settings where laboratory facilities are not well developed, is of key importance in the control of this disease. Rapid diagnostic tests (RDTs) offer great potential to address this need. Reports of significant variation in the field performance of RDTs based on the detection of
Plasmodium falciparum
histidine-rich protein 2 (HRP2) (PfHRP2) and of significant sequence polymorphism in PfHRP2 led us to evaluate the binding of four HRP2-specific monoclonal antibodies (MABs) to parasite proteins from geographically distinct
P. falciparum
isolates, define the epitopes recognized by these MABs, and relate the copy number of the epitopes to MAB reactivity. We observed a significant difference in the reactivity of the same MAB to different isolates and between different MABs tested with single isolates. When the target epitopes of three of the MABs were determined and mapped onto the peptide sequences of the field isolates, significant variability in the frequency of these epitopes was observed. These findings support the role of sequence variation as an explanation for variations in the performance of HRP2-based RDTs and point toward possible approaches to improve their diagnostic sensitivities.
Molecular Dipstick Test for Diagnosis of Sleeping Sickness
Authors: Deborggraeve S. , Claes F. , Laurent T. , Mertens P. , Leclipteux T. , Dujardin J. C. , Herdewijn P. , Büscher P.
Published: August 2006

Human African trypanosomiasis (HAT) or sleeping sickness is a neglected disease that affects poor rural populations across sub-Saharan Africa. Confirmation of diagnosis is based on detection of parasites in either blood or lymph by microscopy. Here we present the development and the first-phase evaluation of a simple and rapid test (HAT-PCR-OC [human African trypanosomiasis-PCR-oligochromatography]) for detection of amplified
Trypanosoma brucei
DNA. PCR products are visualized on a dipstick through hybridization with a gold-conjugated probe (oligochromatography). Visualization is straightforward and takes only 5 min. Controls both for the PCR and for DNA migration are incorporated into the assay. The lower detection limit of the test is 5 fg of pure
T. brucei
DNA. One parasite in 180 μl of blood is still detectable. Sensitivity and specificity for
T. brucei
were calculated at 100% when tested on blood samples from 26 confirmed sleeping sickness patients, 18 negative controls (nonendemic region), and 50 negative control blood samples from an endemic region. HAT-PCR-OC is a promising new tool for diagnosis of sleeping sickness in laboratory settings, and the diagnostic format described here may have wider application for other infectious diseases.
Which sociodemographic factors are important on smoking behaviour of high school students? The contribution of classification and regression tree methodology in a broad epidemiological survey
Authors: Özge C , Toros F , Bayramkaya E , Çamdeviren H , Şaşmaz T
Published: August 2006

Background

The purpose of this study is to evaluate the most important sociodemographic factors on smoking status of high school students using a broad randomised epidemiological survey.

Methods

Using in-class, self administered questionnaire about their sociodemographic variables and smoking behaviour, a representative sample of total 3304 students of preparatory, 9th, 10th, and 11th grades, from 22 randomly selected schools of Mersin, were evaluated and discriminative factors have been determined using appropriate statistics. In addition to binary logistic regression analysis, the study evaluated combined effects of these factors using classification and regression tree methodology, as a new statistical method.

Results

The data showed that 38% of the students reported lifetime smoking and 16.9% of them reported current smoking with a male predominancy and increasing prevalence by age. Second hand smoking was reported at a 74.3% frequency with father predominance (56.6%). The significantly important factors that affect current smoking in these age groups were increased by household size, late birth rank, certain school types, low academic performance, increased second hand smoking, and stress (especially reported as separation from a close friend or because of violence at home). Classification and regression tree methodology showed the importance of some neglected sociodemographic factors with a good classification capacity.

Conclusions

It was concluded that, as closely related with sociocultural factors, smoking was a common problem in this young population, generating important academic and social burden in youth life and with increasing data about this behaviour and using new statistical methods, effective coping strategies could be composed.
Paracetamol availability and recent changes in paracetamol poisoning: is the 1998 legislation limiting availability of paracetamol being followed?
Authors: Greene S L , Dargan P I , Leman P , Jones A L
Published: August 2006

Objective

To determine the degree of adherence to legislation introduced in 1998 restricting the availability of over the counter paracetamol.

Design

A prospective observational study.

Setting

An emergency department in an inner city London teaching hospital. Pharmacy and non-pharmacy outlets in south London.

Main outcome measures

(1) The source of paracetamol ingested by 107 patients presenting with an acute paracetamol overdose (2001–2003) and (2) the ability to purchase paracetamol from pharmacy and non-pharmacy outlets in a manner contravening paracetamol pack size legislation (2004).

Results

Potentially toxic amounts of paracetamol in excess of pack size restrictions were purchased in 70% (17 of 24) of outlets. Forty six per cent of patients who had ingested a potentially toxic dose of paracetamol obtained the tablets in a manner contravening the 1998 legislation.

Conclusion

Legislation limiting the availability of over the counter paracetamol is not being adhered to in south London. A significant number of patients ingesting a potentially toxic dose of paracetamol report purchasing the tablets in a manner contravening the legislation. Studies that attempt to assess the impact of the legislation need to be interpreted in the context of these results. Measures to enforce current legislation may help to reduce the severity of paracetamol poisoning in the UK.
Fournier's gangrene and its emergency management
Authors: Thwaini A , Khan A , Malik A , Cherian J , Barua J , Shergill I , Mammen K
Published: August 2006

Fournier's gangrene (FG) is a rare but life threatening disease. Although originally thought to be an idiopathic process, FG has been shown to have a predilection for patients with diabetes as well as long term alcohol misuse; however, it can also affect patients with non-obvious immune compromise. The nidus is usually located in the genitourinary tract, lower gastrointestinal tract, or skin. FG is a mixed infection caused by both aerobic and anaerobic bacterial flora. The development and progression of the gangrene is often fulminating and can rapidly cause multiple organ failure and death. Because of potential complications, it is important to diagnose the disease process as early as possible Although antibiotics and aggressive debridement have been broadly accepted as the standard treatment, the death rate remains high.
Diagnostic approach to patients with suspected vasculitis
Authors: Suresh E
Published: August 2006

Vasculitis presents several diagnostic challenges. Firstly, patients could present with protean clinical manifestations with a wide spectrum ranging from isolated cutaneous vasculitis to multisystem involvement. Secondly, there are several medical conditions that could mimic the presentation of vasculitis. The range of differential diagnosis is therefore broad. Thirdly, vasculitis could occur as a primary disorder or be secondary to various medical conditions. It becomes important to differentiate them, as treatment of some forms of vasculitis such as those that are secondary to infection or drugs, is different from that of primary vasculitis. Fourthly, there are several different forms of vasculitis. Some are benign and self limiting, while others have the potential to threaten vital organ function and life. It follows that a rational approach is required during evaluation of patients with suspected vasculitis.
Management of urethral strictures
Authors: Mundy A R
Published: August 2006

Controlled clinical trials are unusual in surgery, rare in urology, and almost non-existent as far as the management of urethral stricture is concerned. What data there are come largely from so called “expert opinion” and the quality of this is variable. None the less, the number of so called experts, past and present, is comparatively small and in broad principle their views more or less coincide. Although this review is therefore inevitably biased, it is unlikely that expert opinion will take issue with most of the general points raised here.
Preventing fragility hip fracture in high risk groups: an opportunity missed
Authors: Peng E W K , Elnikety S , Hatrick N C
Published: August 2006

Objective

The national guideline recommends selective case finding as the main strategy by identification of high risk people. This study assessed whether high risk patients were identified before their presentation with fragility fracture.

Methods

A prospective study for 3.5 consecutive months on patients with low energy hip fractures to Brighton and Sussex University Hospital NHS Trusts, which serves a population of 460 000. Data were collected by interview using standardised form, medical record review, and communication with family physicians. Definition of high risk: (1) untreated hypogonadism (2) corticosteroid users (3) disorders with increased bone loss (4) previous fragility fractures.

Results

98 patients were admitted with hip fracture. Thirty nine (40%) had at least one high risk factor. High risk patients (7 of 39, 18%) were no more likely to receive prophylaxis compared with patients without high risk factor (5 of 59, 8%) (p = 0.21). Previous fragility fracture (23) was the commonest risk factor followed by disorders with increased bone loss (10), premature menopause (10), and corticosteroid users (5). Fifteen patients (15%) had susceptibility to frequent falls and two had maternal history of osteoporosis. The proportion of treated patients were 20% (2 of 10) in premature menopause, 10% (1 of 10) in diseases with secondary osteoporosis, 13% (3 of 23) in previous fragility fracture, and 80% (4 of 5) in corticosteroid users (p = 0.01)

Conclusion

Prevention of hip fracture is still inadequate in high risk patients. Discrepancy seemed to exist in treatment frequency among different high risk groups suggesting that emphasis on prevention of osteoporosis has not been reinforced in all people at risk.