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Biomedical subjects

Barbara M Willey

Publications and source records attributed to Barbara M Willey.

14 recordsLinked to original sources
PubMed2006

Severe acute respiratory syndrome diagnostics using a coronavirus protein microarray.

To monitor severe acute respiratory syndrome (SARS) infection, a coronavirus protein microarray that harbors proteins from SARS coronavirus (SARS-CoV) and five additional coronaviruses was constructed. These microarrays were used to screen approximately 400 Canadian sera from the SARS outbreak, including samples from confirmed SARS-CoV cases, respiratory illness patients, and healthcare professionals. A computer algorithm that uses multiple classifiers to predict samples from SARS patients was developed and used to predict 206 sera from Chinese fever patients. The test assigned patients into two distinct groups: those with antibodies to SARS-CoV and those without. The microarray also identified patients with sera reactive against other coronavirus proteins. Our results correlated well with an indirect immunofluorescence test and demonstrated that viral infection can be monitored for many months after infection. We show that protein microarrays can serve as a rapid, sensitive, and simple tool for large-scale identification of viral-specific antibodies in sera.

Algorithms↗
PubMed2006

Verification of the IDI-MRSA assay for detecting methicillin-resistant Staphylococcus aureus in diverse specimen types in a core clinical laboratory setting.

The IDI-MRSA assay has a sensitivity of 96% and a specificity of 96% when used to screen patients at extranasal sites. This verification study used previously unverified swabs and was undertaken in a core medical laboratory using nonmicrobiology technologists trained in sample processing, molecular laboratory work flow, and PCR practice.

Humans↗
PubMed2005

Community-associated methicillin-resistant Staphylococcus aureus in horses and humans who work with horses.

OBJECTIVE: To evaluate the prevalence of nasal colonization with methicillin-resistant Staphylococcus aureus (MRSA) in horses and horse personnel. DESIGN: Prospective prevalence study. SAMPLE POPULATION: 972 horses and 107 personnel from equine farms in Ontario, Canada and New York state. PROCEDURE: Nasal swab specimens were collected from horses and humans on farms with (targeted surveillance) and without (nontargeted surveillance) a history of MRSA colonization or infection in horses during the preceding year. Selective culture for MRSA was performed. Isolates were typed via pulsed-field gel electrophoresis, and antibiograms were determined. RESULTS: MRSA was isolated from 46 of 972 (4.7%) horses (0/581 via nontargeted surveillance and 46/391 [12%] via targeted surveillance). Similarly, MRSA was isolated from 14 of 107 (13%) humans (2/41 [5%] from nontargeted surveillance and 12/66 [18%] from targeted surveillance). All isolates were subtypes of Canadian epidemic MRSA-5, an uncommon strain in humans. All isolates were resistant to at least 1 antimicrobial class in addition to beta-lactams. On all farms with colonized horses, at least 1 human was colonized with an indistinguishable subtype. For horses, residing on a farm that housed > 20 horses was the only factor significantly associated with MRSA colonization. For humans, regular contact with > 20 horses was the only identified risk factor. CONCLUSIONS AND CLINICAL RELEVANCE: Results confirm a reservoir of colonized horses on a variety of farms in Ontario and New York and provide evidence that 1 MRSA strain is predominantly involved in MRSA colonization in horses and humans that work with horses.

Animals↗
PubMed2005

PCR ribotyping of Clostridium difficile isolates originating from human and animal sources.

Molecular typing of Clostridium difficile isolates from animals and humans may be useful for evaluation of the possibility for interspecies transmission. The objective of this study was to evaluate C. difficile isolates from domestic animals and humans using PCR ribotyping. Isolates were also tested using PCR for the presence of genes encoding toxins A and B. One hundred and thirty-three isolates of C. difficile from dogs (n = 92), horses (n = 21) and humans (n = 20), plus one each from a cat and a calf, were evaluated. Overall, 23 ribotypes were identified. Of these, nine were identified from dogs, 12 from horses, seven from humans and one each from the cat and calf. In dogs, humans and horses, one or two different ribotypes predominated. Overall, 25 % of isolates from humans were indistinguishable from isolates from one or more animal species. Genes encoding C. difficile toxins A and B were detected in all human, equine and bovine isolates, and in 69 % of canine isolates. While different ribotypes appear to predominate in different mammalian species, several indistinguishable strains may be found in multiple species. This suggests that there is a potential for interspecies transmission of C. difficile and epidemiological studies are warranted.

Animals↗
PubMed2005

Identification of a progenitor of the CTX-M-9 group of extended-spectrum beta-lactamases from Kluyvera georgiana isolated in Guyana.

Chromosomal beta-lactamase genes (bla(KLUY)) from six Kluyvera georgiana strains isolated in Guyana were cloned and expressed in Escherichia coli. KLUY-1 exhibited 100% amino acid identity with the extended-spectrum beta-lactamase CTX-M-14. We also show that a 2.7-kb Kluyvera chromosomal region exhibits 99% nucleotide identity to a portion of In60 that includes bla(CTX-M-9).

Amino Acid Sequence↗
PubMed2005

Use of a selective enrichment broth to recover Clostridium difficile from stool swabs stored under different conditions.

The recovery of Clostridium difficile from the stools of patients with C. difficile-associated diarrhea was evaluated by use of an enrichment broth (cycloserine-cefoxitin fructose broth supplemented with 0.1% sodium taurocholate [TCCFB]) and was compared to that from selective agar (cycloserine-cefoxitin fructose agar [CCFA]) and alcohol shock followed by inoculation onto blood agar (AS-BA). TCCFB was superior to CCFA and AS-BA, and neither the storage time nor the storage temperature affected the recovery rate.

Bacteriological Techniques↗
PubMed2004

Fatal severe acute respiratory syndrome is associated with multiorgan involvement by coronavirus.

Severe acute respiratory syndrome (SARS) is characterized by pulmonary compromise; however, patients often have evidence of other organ dysfunction that may reflect extrapulmonary dissemination of SARS coronavirus (SARS-CoV). We report on the distribution and viral load of SARS-CoV in multiple organ samples from patients who died of SARS during the Toronto outbreak. SARS-CoV was detected in lung (100%), bowel (73%), liver (41%), and kidney (38%) in 19 patients who died of SARS, with the highest viral loads observed in lung (1.0 x 10(10) copies/g) and bowel (2.7 x 10(9) copies/g). Fatal SARS was associated with multiorgan viral dissemination in a distribution that has implications for disease manifestation, viral shedding, and transmission.

Coronavirus↗
PubMed2004

Complete nucleotide sequence of a 92-kilobase plasmid harboring the CTX-M-15 extended-spectrum beta-lactamase involved in an outbreak in long-term-care facilities in Toronto, Canada.

A major outbreak involving an Escherichia coli strain that was resistant to expanded-spectrum cephalosporins occurred in Toronto and surrounding regions in 2000 to 2002. We report the complete sequence of a plasmid, pC15-1a, that was found associated with the outbreak strain. Plasmid pC15-1a is a circular molecule of 92,353 bp consisting of two distinct regions. The first is a 64-kb region that is essentially homologous to the non-R-determinant region of plasmid R100 except for several point mutations, a few small insertions and deletions, and the absence of Tn10. The second is a 28.4-kb multidrug resistance region (MDR) that has replaced the R-determinant region of the R100 progenitor and consists mostly of transposons or partial transposons and five copies of the insertion element IS26. All drug resistance genes found in pC15-1a, including the beta-lactamase genes bla(CTX-M-15), bla(OXA-1), and bla(TEM-1), the tetracycline resistance gene tetA, and aminoglycoside resistance genes aac(6')-Ib and aac(3)-II, are located in the MDR. The bla(CTX-M-15) gene was found downstream of ISEcp1as part of a transposition unit, as determined from the surrounding sequence. Examination of the plasmids from CTX-M-15-harboring strains isolated from hospitals across Canada showed that pC15-1a was found in several strains isolated from a site in western Canada. Comparison of pC15-1a and pCTX15, found in an E. coli strain isolated in India in 1999, revealed that the plasmids had several features in common, including an R100 backbone and several of the resistance genes, including bla(CTX-M-15), bla(TEM-1), bla(OXA-1), tetA, and aac(6')-Ib.

Base Sequence↗
PubMed2004

Use of the Vitek-1 and Vitek-2 systems for detection of constitutive and inducible macrolide resistance in group B streptococci.

A prospective study of erythromycin and clindamycin resistance was performed with 304 consecutive group B streptococci (GBS) isolates. According to two automated susceptibility testing systems, Vitek-1 and Vitek-2, and double-disk agar diffusion, 79.9% were susceptible to both erythromycin and clindamycin. However, for macrolide-lincosamide-streptogramin B-inducible isolates, the accuracies of the Vitek-1 and Vitek-2 systems were 5.6 and 94.4%. In light of these results, we recommend that GBS be routinely tested using the double-disk diffusion method.

Clindamycin↗
PubMed2004

Severe acute respiratory syndrome-associated coronavirus in lung tissue.

Efforts to contain severe acute respiratory syndrome (SARS) have been limited by the lack of a standardized, sensitive, and specific test for SARS-associated coronavirus (CoV). We used a standardized reverse transcription-polymerase chain reaction assay to detect SARS-CoV in lung samples obtained from well-characterized patients who died of SARS and from those who died of other reasons. SARS-CoV was detected in all 22 postmortem lung tissues (to 10(9) viral copies/g) from 11 patients with probable SARS but was not detected in any of the 23 lung control samples (sample analysis was blinded). The sensitivity and specificity (95% confidence interval) were 100% (84.6% to 100%) and 100% (85.1% to 100%), respectively. Viral loads were significantly associated with a shorter course of illness but not with the use of ribavirin or steroids. CoV was consistently identified in the lungs of all patients who died of SARS but not in control patients, supporting a primary role for CoV in deaths.

Adult↗
PubMed2003

Emergence of levofloxacin-resistant pneumococci in immunocompromised adults after therapy for community-acquired pneumonia.

We describe 4 patients infected with levofloxacin-resistant pneumococci after therapy for community-acquired pneumonia (CAP). The 4 patients had 15 episodes of CAP; Streptococcus pneumoniae was isolated from blood or sputum samples obtained during 14 of the episodes. The underlying medical condition was Bruton agammaglobulinemia in 3 patients and chronic lymphoid leukemia in the other. The initial episode of CAP in each patient was due to a levofloxacin-susceptible strain. One of 4 reinfections and 5 of 6 relapses were due to levofloxacin-resistant strains. All of these strains had amino acid substitutions in the quinolone-resistance-determining region of the genes parC and gyrA. The time between episodes of pneumonia varied from 1 to 4 months. In immunocompromised patients with suspected or proven pneumococcal infection, it may be prudent not to use fluoroquinolone monotherapy empirically when the patient has a history of fluoroquinolone therapy in at least the past 4 months.

Adult↗
PubMed2003

Emergence of macrolide resistance in throat culture isolates of group a streptococci in Ontario, Canada, in 2001.

Of 500 group A streptococci isolated from pharyngeal swabs, 72 (14.4%) were macrolide resistant, compared to 2.1% in 1997. Of these, 66 (92%) were of the M phenotype and 6 (8.3%) were of the MLS phenotype. Pulsed-field gel electrophoresis found that two clones, with patterns identical to those of serotypes M1 and M4, accounted for 19.4 and 68.1% of the macrolide-resistant isolates, respectively.

Anti-Bacterial Agents↗
PubMed2003

Investigation of Staphylococcus aureus isolates identified as erythromycin intermediate by the Vitek-1 System: comparison with results obtained with the Vitek-2 and Phoenix systems.

We identified 69 Staphylococcus aureus isolates that were erythromycin intermediate as reported by the Vitek-1 system using the GPS-105 card. Of the 57 strains that were available for further testing, all were erythromycin resistant by broth microdilution and the Phoenix system, while the Vitek-2 system identified 55 of 57 strains (96%) as erythromycin resistant. The majority of isolates (54 of 57 [95%]) exhibited the inducible MLS (macrolide-lincosamide-streptogramin family) phenotype, as shown by the double-disk test. We recommend that all S. aureus strains determined as erythromycin intermediate by the Vitek-1 system be interpreted as resistant to erythromycin.

Anti-Bacterial Agents↗