bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of the PACE 2 Assay, Two Amplification Assays, and Clearview EIA for Detection of
Chlamydia trachomatis
in Female Endocervical and Urine Specimens
Authors: Lauderdale Tsai-Ling , Landers Lenore , Thorneycroft Ian , Chapin Kimberle 
Published: December 2019

ABSTRACT

Screening for sexually transmitted diseases (STDs) in a greater proportion of sexually active patients has become an accepted protocol by most health care providers. The purpose of this study was to compare the current test methods for detection of
Chlamydia trachomatis
used at the University of South Alabama, the PACE 2 assay (Gen-Probe) and the Clearview EIA (Wampole Laboratories), with two amplification technologies, the AMP CT (Gen-Probe) and LCx (Abbott) assays. In addition, a number of demographic parameters were ascertained by asking questions at the time of examination as well as for health care provider concerns and preferences. One urine and four endocervical swab specimens were collected in random order from 787 female patients attending one of four obstetrics-gynecology clinics. Eighty-seven percent of patients had no STD-related symptoms. Patients were considered positive for
C. trachomatis
if three or more assays (swab and/or urine) were positive. Abbott and Gen-Probe confirmed discrepant results by alternate amplified assays. A total of 66 true-positive specimens were detected by use of the combination of endocervical swabs and urine specimens. After discrepant analysis, sensitivities for endocervical swab specimens for the EIA and the PACE 2, LCx, and AMP CT assays were 50, 81, 97, and 100%, respectively. Sensitivities for the LCx and AMP CT assays with urine specimens were 98 and 81%, respectively. The prevalence of
C. trachomatis
was 8.4%, as determined by amplification technology. Overall, the amplification technologies were the most sensitive methods with either swab (AMP CT assay) or urine (LCx assay) specimens. The PACE 2 assay offered the advantage of a simpler and less expensive assay with acceptable sensitivity. The clearview CT EIA, while yielding a rapid in-office result, had unacceptably low sensitivity. The wide variation in performance with amplification assays with urine specimens as reported in both this study and the literature obviates the need to clarify optimal parameters for this specimen type.
Usefulness of Fatty Acid Composition for Differentiation of
Legionella
Species
Authors: Diogo Alexandra , Veríssimo António , Nobre M. Fernanda , da Costa Milton S. 
Published: December 2019

ABSTRACT

Numerical analysis of fatty acid methyl ester (FAME) profiles of 199 isolates and 76 reference strains, belonging to all validly described species of the genus
Legionella
that can be cultured in laboratory media, was used to differentiate between the species of this genus. With the exception of the strains that autofluoresced red, it was possible to differentiate all the other
Legionella
species. The strains of the species
L. bozemanii
,
L. dumoffii
,
L. feeleii
,
L. gormanii
,
L. maceachernii
,
L. micdadei
, and
L. quinlivanii
did not form single clusters, showing some degree of variability in the fatty acid compositions. The strains of the blue-white autofluorescent species had very similar fatty acid compositions and were difficult to distinguish from each other. Nine isolates had fatty acid profiles unlike those of any of the validly described species and may represent different FAME groups of known species or undescribed
Legionella
species. The method used in this study was useful for screening and discriminating large number of isolates of
Legionella
species. Moreover, the results obtained can be included in a database of fatty acid profiles, leading to a more accurate automatic identification of
Legionella
isolates.
Rapid Detection of Respiratory Viruses by Shell Vial Assay Using Simultaneous Culture of HEp-2, LLC-MK2, and MDCK Cells in a Single Vial
Authors: Navarro-Marí José M. , Sanbonmatsu-Gámez Sara , Pérez-Ruiz Mercedes , De La Rosa-Fraile Manuel 
Published: December 2019

ABSTRACT
A shell vial assay with simultaneous culture of HEp-2, LLC-MK2, and MDCK cell lines in a single tube (CoHLM SV assay) was compared with traditional tube culture (TC) for the detection of the main respiratory viruses in 358 nasal wash specimens. A total of 170 strains were isolated from 168 virus-positive samples. A total of 94.1% of the strains (160 strains; 128 respiratory syncytial viruses and 32 other viruses) were detected by the CoHLM SV assay in 48 h, whereas 98.2% of the strains (167 strains; 132 respiratory syncytial viruses and 35 other viruses) were detected by TC in a mean time of 6 days. The CoHLM SV assay may be useful for the rapid detection of respiratory viruses.
Inactivation of
Mycobacterium tuberculosis
for DNA Typing Analysis
Authors: Bemer-Melchior P. , Drugeon H. B. 
Published: December 2019

ABSTRACT

DNA fingerprinting analysis of
Mycobacterium tuberculosis
is used for epidemiological studies and the control of laboratory cross-contamination. Because standardized procedures are not entirely safe for mycobacteriology laboratory staff, the paper proposes a new technique for the processing of specimens. The technique ensures the inactivation of
M. tuberculosis
before DNA extraction without the loss of DNA integrity. The control of inactivated cultures should be rigorous and should involve the use of two different culture media incubated for at least 4 months.
Prototypal Diarrheagenic Strains of
Hafnia alvei
Are Actually Members of the Genus
Escherichia
Authors: Janda J. Michael , Abbott Sharon L. , Albert M. John 
Published: December 2019

ABSTRACT

We analyzed five bacterial strains, designated 19982, 9194, 10457, 10790, and 12502, that were isolated from stool specimens of individuals with diarrheal illness by the International Centre for Diarrhoeal Disease Research in Dhaka, Bangladesh (M. J. Albert, S. M. Faruque, M. Ansaruzzaman, M. M. Islam, K. Haider, K. Alam, I. Kabir, and R. Robins-Browne, J. Med. Microbiol. 37:310–314, 1992). The strains were initially identified as
Hafnia alvei
with a commercial identification system and were reported to contain the
eae
gene of enteropathogenic
Escherichia coli
. Results of conventional biochemical analyses, testing of susceptibility to cephalothin, lysis by a
Hafnia
-specific phage, and amplification of the outer membrane protein gene
phoE
with species-specific primers support the identification of these strains as members of the genus
Escherichia
rather than
Hafnia alvei
. These strains varied from typical
E. coli
strains by their inability to produce acid from lactose or
d
-sorbitol and failure to elaborate the enzyme β-
d
-glucuronidase. PCR analysis confirmed previous findings that the strains were positive for the
eae
gene and negative for other virulence markers present among recognized categories of diarrheagenic
E. coli
. Our findings support the hypothesis that these strains are a new category of diarrheagenic isolates belonging to the genus
Escherichia
and illustrate the importance of using multiple methodologies when identifying new bacterial agents of diarrheal disease.
Involvement of Enterotoxins G and I in Staphylococcal Toxic Shock Syndrome and Staphylococcal Scarlet Fever
Authors: Jarraud Sophie , Cozon Grégoire , Vandenesch François , Bes Michèle , Etienne Jerome , Lina Gerard 
Published: December 2019

ABSTRACT

We investigated the involvement of the recently described staphylococcal enterotoxins G and I in toxic shock syndrome. We reexamined
Staphylococcus aureus
strains isolated from patients with menstrual and nonmenstrual toxic shock syndrome (nine cases) or staphylococcal scarlet fever (three cases). These strains were selected because they produced none of the toxins known to be involved in these syndromes (toxic shock syndrome toxin 1 and enterotoxins A, B, C, and D), enterotoxin E or H, or exfoliative toxin A or B, despite the fact that superantigenic toxins were detected in a CD69-specific flow cytometry assay measuring T-cell activation. Sets of primers specific to the enterotoxin G and I genes (
seg
and
sei
, respectively) were designed and used for PCR amplification. All of the strains were positive for
seg
and
sei
. Sequence analysis confirmed that the PCR products, corresponded to the target genes. We suggest that staphylococcal enterotoxins G and I may be capable of causing human staphylococcal toxic shock syndrome and staphylococcal scarlet fever.
Comparison of the Second-Generation Digene Hybrid Capture Assay with the Branched-DNA Assay for Measurement of Hepatitis B Virus DNA in Serum
Authors: Ho Stephen K. N. , Chan Tak Mao , Cheng Ignatius K. P. , Lai Kar Neng 
Published: December 2019

ABSTRACT

The optimal hepatitis B virus (HBV) DNA quantitative assay for clinical use remains to be determined. We examined the sensitivity, linearity, and variability of a novel second-generation antibody capture solution hybridization assay, the Digene Hybrid Capture II assay (HCII), and compared it with another widely used solution hybridization assay, the branched-DNA (bDNA) assay (Quantiplex; Chiron Corp.). Our results showed similar and satisfactory assay linearity values, as well as interassay and intra-assay variability values, for both HCII and bDNA assays across different ranges of HBV DNA. Ninety-one percent of 102 serum samples from hepatitis B surface antigen-positive patients showed concordant results with the two assays. The HCII assay was more sensitive than the bDNA assay by 1 dilution, with the lowest reading being 0.9 pg/ml (3.8 pg/ml by bDNA assay). The HBV DNA seropositivity rates for the 102 samples were 58, 67, and 97% by bDNA, HCII, and nested PCR, respectively. While the relationship between results obtained with the bDNA assay and those with the HCII assay was nonlinear, with the bDNA assay yielding values 2.83 ± 0.92-fold higher than those of the HCII assay, especially at high HBV DNA levels, a linear relationship was observed between the two sets of data after logarithmic conversion. The formula for interassay conversion of results was derived as follows: HBV DNA by HCII (picograms per milliliter) = 3.19 × [HBV DNA by bDNA (megaequivalents per milliliter)]
0.866
. The HCII assay was technically less complex and required a shorter assay time (4 h) than the bDNA assay (24 h). We conclude that the HCII assay compares favorably with the bDNA assay and offers the additional advantages of increased sensitivity and shorter assay time. The increased sensitivity should be particularly useful in monitoring the efficacy of antiviral therapies and detecting the emergence of drug-resistant HBV mutants.
Value of Enterobacterial Repetitive Intergenic Consensus PCR for Study of
Pasteurella multocida
Strains Isolated from Mouths of Dogs
Authors: Loubinoux Julien , Lozniewski Alain , Lion Christine , Garin Daniel , Weber Michèle , Le Faou Alain E. 
Published: December 2019

ABSTRACT

Fifty-six
Pasteurella multocida
strains (40
P. multocida
subsp.
septica
and 16
P. multocida
subsp.
multocida
strains) isolated from the mouths of 56 dogs among the 134 living in a French canine military training center (132e Groupe Cynophile de l’Armée de Terre, Suippes, France) were studied by use of enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) and restriction fragment length polymorphism (RFLP) techniques. Both techniques showed genomic heterogeneity of the strains studied. However, RFLP was more discriminatory than ERIC-PCR for differentiating
P. multocida
strains. All but three pairs of strains were discriminated by RFLP, suggesting a limited circulation of strains between these dogs living in proximity. Although ERIC-PCR is easier and faster to perform, it cannot be recommended for epidemiological studies of
P. multocida
strains.
Phialemonium
Fungemia: Two Documented Nosocomial Cases
Authors: Guarro Josep , Nucci Marcio , Akiti Tiyomi , Gené Josepa , Cano Josep , Barreiro M. Da Gloria C. , Aguilar Carme 
Published: December 2019

ABSTRACT

Two fungal isolates recovered from the blood of two immunosuppressed patients are described as
Phialemonium curvatum
. One patient died, while the other, who was infected with
Exophiala jeanselmei
at the same time, survived after successful treatment with itraconazole. Analysis of internal transcribed spacer sequences demonstrated that the isolates belonged to the same strain and that the source of infection was probably a catheter. The taxonomic position of
P. curvatum
is discussed, and
Phialemonium dimorphosporum
is considered a synonym. The in vitro inhibitory activities of six antifungal agents (amphotericin B, itraconazole, ketaconazole, miconazole, flucytosine, and fluconazole) were determined against seven isolates of
Phialemonium
. Except for flucytosine, all of them were remarkably effective.
Phialemonium
should be added to the list of potential causes of nosocomial fungemia in cancer patients.
Molecular Epidemiological Study ofHaemophilus influenzaeSerotype b Strains Obtained from Children with Meningitis in Japan
Authors: Mitsuda Toshihiro , Kuroki Haruo , Ishikawa Nobuyasu , Imagawa Tomoyuki , Ito Schuichi , Miyamae Takako , Mori Masaaki , Uehara Suzuko , Yokota Shumpei 
Published: December 2019

ABSTRACTWe report an epidemiological study of 30Haemophilus influenzaeserotype b (Hib) strains derived from the cerebrospinal fluid of children with meningitis. The Hib strains were biotyped, tested for β-lactamase production, and genotyped by long PCR-ribotyping, random amplified polymorphic DNA (RAPD) analysis, and genomic DNA restriction fragment length polymorphism (RFLP) analysis by pulsed-field gel electrophoresis (PFGE). The phenotypic study characterized 22 of the strains (73%) as biotype I. A genotypic study using long PCR-ribotyping withHaeIII restriction digestion showed no polymorphisms among these 30 Hib strains, but RAPD analysis with two sets of primers demonstrated two distinctive subtypes: one typical of the strains of biotype group II and the second characteristic of the strains of biotype groups I and IV. Each RAPD group was subtyped into several genotypic groups by PFGE-RFLP withSmaI digestion. The genotyping of clinically isolated Hib strains may help to elucidate transmission routes in community infections, endemicity, and the reasons for vaccine failure.
Usefulness of Spoligotyping To Discriminate IS
6110
Low-Copy-Number
Mycobacterium tuberculosis
Complex Strains Cultured in Denmark
Authors: Bauer Jeanett , Andersen Åse B. , Kremer Kristin , Miörner Håkan 
Published: December 2019

ABSTRACT

Mycobacterium tuberculosis
complex strains cultured in Denmark have been analyzed by IS
6110
restriction fragment length polymorphism (RFLP) on a routine basis from 1992 and onwards. Due to the influx of immigrants with tuberculosis, the number of strains harboring only one to five copies of IS
6110
has increased steadily. Since the discriminatory power of IS
6110
fingerprinting for such strains is poor, we have performed additional genotyping of all low-copy-number strains by the recently described PCR-based method known as spoligotyping. A total of 311 clinical strains were typed: 14
Mycobacterium bovis
BCG, 48
M. bovis
, and 249
M. tuberculosis
strains. Spoligotyping correctly differentiated
M. bovis
and
M. bovis
BCG from
M. tuberculosis
strains, but it did not differentiate
M. bovis
from
M. bovis
BCG. All
M. bovis
BCG strains exhibited identical spoligotype patterns. The discriminatory power of spoligotyping of low-copy-number
M. tuberculosis
strains was higher than that of IS
6110
fingerprinting. Based on RFLP typing solely, 83% of the low-copy-number
M. tuberculosis
strains were found to form part of a cluster, and 75% were found to form a cluster on the basis of spoligotyping. When the two techniques were combined, the amount of clustering decreased to 55%. The combination of these two techniques might be valuable in studying the epidemiology of
M. tuberculosis
strains harboring few copies of the IS
6110
element.
Improved Detection of Hepatitis B Virus Surface Antigen by a New Rapid Automated Assay
Authors: Weber Bernard , Bayer Anja , Kirch Peter , Schlüter Volker , Schlieper Dietmar , Melchior Walter 
Published: December 2019

ABSTRACT
The performance of hepatitis B virus (HBV) surface antigen (HBsAg) screening assays is continuously improved in order to reduce the residual risk of transfusion-associated hepatitis B. In a multicenter study, a new automated rapid screening assay, Elecsys HBsAg (Roche Diagnostics), was compared to well-established tests (Auszyme Monoclonal [overnight incubation] version B and IMx HBsAg [Abbott]). Included in the evaluation were 23 seroconversion panels; sera from the acute and chronic phases of infection; dilution series of various HBsAg standards, HBV subtypes, and S gene mutants; and isolated anti-HBV core antigen-positive samples. To challenge the specificity of the new assay, sera from HBsAg-negative blood donors, pregnant women, and dialysis and hospitalized patients and potentially cross-reactive samples were investigated. Elecsys HBsAg showed a higher sensitivity for HBsAg subtypes ad, ay, adw2, adw4, ayw1, ayw2, ayw4, and adr detection in dilution series of different standards or sera than Auszyme Monoclonal version B and/or IMx HBsAg. Acute hepatitis B was detected in 11 to 16 of 23 seroconversion panels between 2 and 16 days earlier with Elecsys HBsAg than with the alternative assays. Elecsys HBsAg and Auszyme Monoclonal version B detected HBsAg surface mutants with equal sensitivity. The sensitivity and specificity of Elecsys HBsAg were 100%. Auszyme Monoclonal version B had a 99.9% specificity, and its sensitivity was 96.6%. IMx HBsAg showed a poorer sensitivity and specificity than the other assays. In conclusion, Elecsys HBsAg permits earlier detection of acute hepatitis B and different HBV subtypes than the alternative assays. By using highly sensitive HBsAg screening assays, low-level HBsAg carriers among isolated anti-HBV core antigen-positive individuals can be detected.
Mutations in the
rpoB
Gene of Rifampin-Resistant
Mycobacterium tuberculosis
Strains Isolated Mostly in Asian Countries and Their Rapid Detection by Line Probe Assay
Authors: Hirano Kazue , Abe Chiyoji , Takahashi Mitsuyoshi 
Published: December 2019

ABSTRACT

Mutations in the
rpoB
gene of 90 rifampin-resistant
Mycobacterium tuberculosis
isolates mostly from Asian countries were analyzed. Ten distinct single-nucleotide substitutions were found among the isolates by automated sequencing. A 3-nucleotide insertion was found in two isolates, and no mutation was found in five isolates (5.6%). A reverse hybridization-based line probe assay (INNO-LiPA Rif TB) for rapid detection of the mutations was evaluated with these isolates. Concordance rates with sequencing results for five wild-type probes (S probes) and four probes for specific mutations (R probes) were 96.7 and 100%, respectively. The overall concordance rate with the in vitro susceptibility testing results was 92.2% (83 of 90 isolates). These results indicate that a commercial line probe assay kit may be useful for rapid diagnosis of rifampin-resistant tuberculosis.
Detection of
Staphylococcus aureus
and
Staphylococcus epidermidis
in Clinical Samples by 16S rRNA-Directed In Situ Hybridization
Authors: Krimmer Vanessa , Merkert Hilde , von Eiff Christof , Frosch Matthias , Eulert Jochen , Löhr Jochen F. , Hacker Jörg , Ziebuhr Wilma 
Published: December 2019

ABSTRACT

Staphylococcus epidermidis
and
Staphylococcus aureus
are the most common causes of medical device-associated infections, including septicemic loosenings of orthopedic implants. Frequently, the microbiological diagnosis of these infections remains ambiguous, since at least some staphylococci have the capacity to reduce their growth rate considerably. These strains exhibit a small-colony phenotype, and often they are not detectable by conventional microbiological techniques. Moreover, clinical isolates of
S. aureus
and
S. epidermidis
adhere to polymer and metal surfaces by the generation of thick, multilayered biofilms consisting of bacteria and extracellular polysaccharides. This study reports improved detection and identification of
S. aureus
and
S. epidermidis
by an in situ hybridization method with fluorescence-labeled oligonucleotide probes specific for staphylococcal 16S rRNA. The technique has proven to be suitable for the in situ detection of staphylococci, which is illustrated by the identification of
S. epidermidis
in a connective tissue sample obtained from a patient with septicemic loosening of a hip arthroplasty. We also show that this technique allows the detection of intracellularly persisting bacteria, including small-colony variants of
S. aureus
, and the differentiation of
S. epidermidis
from other clinically relevant staphylococci even when they are embedded in biofilms. These results suggest that the 16S rRNA in situ hybridization technique could represent a powerful diagnostic tool for the detection and differentiation of many other fastidious microorganisms.
Use of Granada Medium To Detect Group B Streptococcal Colonization in Pregnant Women
Authors: Rosa-Fraile Manuel , Rodriguez-Granger Javier , Cueto-Lopez Marina , Sampedro Antonio , Gaye Enrique Biel , Haro José Manuel , Andreu Antonia 
Published: December 2019

ABSTRACT
Direct inoculation onto Granada medium (GM) in plates and tubes was compared to inoculation into a selective Todd-Hewitt broth (with 8 μg of gentamicin per ml and 15 μg of nalidixic acid per ml) for detection of group B streptococci (GBS) in pregnant women with 800 vaginal and 450 vaginoanorectal samples. Comparatively, GM was found to be as sensitive as the selective broth for the detection of GBS in vaginal specimens and more sensitive than selective broth for the detection of GBS in vaginoanorectal samples (96 versus 82%). The use of GM improved the time to reporting of a GBS-positive result by at least 24 h and reduced the direct cost of screening. We have also found that the inconvenience of anaerobic incubation of GM plates can be avoided when a cover slide is placed upon the inoculum, because aerobic incubation in GM plates with cover slides causes GBS to develop the same pigmentation that it develops with incubation under anaerobic conditions. These data support the routine use of GM plates or tubes as a more accurate, easier, and cheaper method of identification of GBS-colonized women compared to the enrichment broth technique.
Versatile Fluorescent Staining of Fungi in Clinical Specimens by Using the Optical Brightener Blankophor
Authors: Rüchel R. , Schaffrinski Meike 
Published: December 2019

ABSTRACT
Fluorescent staining of fungi in clinical specimens with the optical brightener Blankophor can be performed concomitantly with maceration of surrounding tissue and may be accelerated by heating. The procedure is suitable for disclosing fungi in gram-stained microscopical mounts and can be used for screening of tissue sections prior to immunofluorescence.