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Biomedical subjects

Kurt D Reed

Publications and source records attributed to Kurt D Reed.

At least 19 recordsLinked to original sources

A LightCycler TaqMan assay for detection of Borrelia burgdorferi sensu lato in clinical samples.

Lyme disease (LD) is an infection caused by an ixodid tick-borne spirochete, Borrelia burgdorferi sensu lato. LD manifests itself as a multisystem inflammatory disease that affects the skin in its early localized stage and spreads to the joints, nervous system, heart, and, to a lesser extent, other organ systems in its later disseminated stages. If diagnosed and treated early with appropriate antibiotics, LD is almost always readily cured. Developing a highly sensitive and specific real-time polymerase chain reaction assay could be very useful in improving the diagnostic accuracy and decreasing turnaround time for results. We report the development of a LightCycler TaqMan assay targeting the OspA gene for clinical detection of B. burgdorferi sensu lato in various types of biologic samples. This assay was validated by testing a variety of clinical samples including cerebrospinal fluid, synovial fluid, skin biopsies, and blood and culture isolates from skin biopsies. The TaqMan testing results were 100% concordant with previously reported results. Reference strains representing isolates from other geographic regions were also successfully amplified. The developed assay is robust, is highly sensitive and specific for B. burgdorferi sensu lato, and is suitable for clinical detection of the bacterium in biologic samples.

Animals↗

Coinfections acquired from ixodes ticks.

The pathogens that cause Lyme disease (LD), human anaplasmosis, and babesiosis can coexist in Ixodes ticks and cause human coinfections. Although the risk of human coinfection differs by geographic location, the true prevalence of coinfecting pathogens among Ixodes ticks remains largely unknown for the majority of geographic locations. The prevalence of dually infected Ixodes ticks appears highest among ticks from regions of North America and Europe where LD is endemic, with reported prevalences of < or =28%. In North America and Europe, the majority of tick-borne coinfections occur among humans with diagnosed LD. Humans coinfected with LD and babesiosis appear to have more intense, prolonged symptoms than those with LD alone. Coinfected persons can also manifest diverse, influenza-like symptoms, and abnormal laboratory test results are frequently observed. Coinfecting pathogens might alter the efficiency of transmission, cause cooperative or competitive pathogen interactions, and alter disease severity among hosts. No prospective studies to assess the immunologic effects of coinfection among humans have been conducted, but animal models demonstrate that certain coinfections can modulate the immune response. Clinicians should consider the likelihood of coinfection when pursuing laboratory testing or selecting therapy for patients with tick-borne illness.

Animals↗

Dissecting plague.

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Disease Outbreaks↗

Bilateral prosthetic knee infection by Campylobacter fetus.

We present the first documented case of a bilateral prosthetic knee joint infection with Campylobacter fetus. Our patient's risk factors included age, the presence of prosthetic joints, and potential exposure through his contact with farm animals. It is important to be aware of the possibility of C fetus joint infections in high-risk patients who present with pain after total joint arthroplasty.

Aged↗

Identification of mosquito bloodmeal source by terminal restriction fragment length polymorphism profile analysis of the cytochrome B gene.

Previous studies have shown that polymerase chain reaction (PCR) heteroduplex analysis (HDA) of the cytochrome B (cytb) gene is useful in identifying mosquito bloodmeals derived from avian hosts. However, interpretation of PCR-HDA gels is performed visually, which can make it difficult to analyze large numbers of specimens and to compare results between laboratories. We investigated the utility of a terminal restriction fragment length polymorphism (T-RFLP) assay to analyze cytb PCR products. PCR was performed on 123 blood or tissue samples from 55 avian, 13 mammalian, and one amphibian species by using end-labeled primers to amplify a 358-bp segment of cytb. Each PCR product was sequenced to determine predicted terminal restriction fragment (TRF) profiles. Additionally, experimental TRFs were determined by sizing fragments from restriction endonuclease digests with capillary electrophoresis. A Web-based searchable database was created to compare unknown mosquito bloodmeal TRF profiles against sequence-predicted and experimentally derived terminal fragment lengths of known vertebrates. The predictive value of experimental profiles was found to be accurate to the species level for 67 of 69 species (97%). Fifty-nine field-collected mosquitoes were tested to determine the bloodmeal source using the T-RFLP method. The bloodmeal source from 50 of these mosquitoes was identified by comparing the TRF profile of the unknown source against the cytochrome B database. The bloodmeal source from the remaining nine mosquitoes was not identified as no known profile matched the experimentally derived profile. T-RFLP analysis is a highly reproducible technique and the searchable TRF database is continually being expanded to include additional species from diverse geographic areas.

Animals↗

Serologic evidence of West Nile virus infection in three wild raptor populations.

We assayed for West Nile virus (WNV) antibodies to determine the presence and prevalence of WNV infection in three raptor populations in southeast Wisconsin during 2003-04. This study was conducted in the framework of ongoing population studies that started before WNV was introduced to the study area. For 354 samples, 88% of 42 adult Cooper's hawks (Accipiter cooperii), 2.1% of 96 nestling Cooper's hawks, 9.2% of 141 nestling red-tailed hawks (Buteo jamaicensis), and 12% of 73 nestling great horned owls (Bubo virginianus) tested positive for WNV antibodies by the constant virus-serum dilution neutralization test. Samples that tested positive for WNV antibodies were collected across a wide variety of habitat types, including urban habitats (both high and low density), roads, parking areas, recreational areas, croplands, pastures, grasslands, woodlands, and wetlands. Based on the increased prevalence and significantly higher WNV antibody titers in adults compared with nestlings, we suggest that nestlings with detectable antibody levels acquired these antibodies through passive transmission from the mother during egg production. Low levels of WNV antibodies in nestlings could serve as a surrogate marker of exposure in adult raptor populations. Based on breeding population densities and reproductive success over the past 15 yr, we found no apparent adverse effects of WNV infections on these wild raptor populations.

Animals↗

Surveillance of above- and below-ground mosquito breeding habitats in a rural midwestern community: baseline data for larvicidal control measures against West Nile Virus vectors.

BACKGROUND: Mosquitoes in the genus Culex are thought to play a major role as vectors in the transmission cycle of West Nile virus (WNV) and other arboviruses in the United States. Effective control of mosquitoes through larviciding and adulticiding is expensive for communities and should be guided by reliable surveillance data on the distribution of mosquito breeding habitats. However, few small to medium sized cities in rural areas of the midwestern United States have this type of baseline information available. OBJECTIVE: During the summer of 2004, we investigated the characteristics of Culex and other mosquito-breeding habitats in a rural central Wisconsin community with a population of approximately 19,000. Such baseline information will aid in the development of rational strategies to control mosquito populations and prevent human exposure to WNV and other mosquito-transmitted viruses. METHODS: Mosquito larvae were collected and identified weekly from 14 below-ground storm water catch basins and 10 above-ground standing water sites distributed throughout the community. Collection began June 4, 2004 and continued through September 24, 2004. For each collection site the primary and adjacent land use patterns were determined. RESULTS: Over the study period, 1,244 larvae were collected from catch basins; 94% were Culex species. Breeding activity was first detected in early July. Peak breeding was observed during a period of several weeks when average daily temperatures were at the maximum observed and rainfall had declined. Organically enriched catch basins in low intensity urban sites adjacent to forests and wetlands were found to be more productive breeding habitats compared to catch basins having little organic debris located in isolated high intensity urban sites. Above-ground standing water sites produced 1,504 larvae; 66% of which were Culex species. Flood control ditches and permanent wetlands with stagnant water were most productive, while ditches with moving water were least productive habitats. Larvae were produced earlier in the season by above-ground sites than were produced by catch basins. However, larvae production was more variable in above-ground sites since half the sites became dry at some point during the study period. CONCLUSION: The observed differences in Culex larvae production based on the variables of habitat-type, temperature, and precipitation support the need for ongoing surveillance in communities to guide public health officials in planning for and prioritizing mosquito control efforts.

Animals↗

Efficacy of the influenza vaccine in patients with malignant lymphoma.

BACKGROUND: The benefits and efficacy of the influenza vaccine have been controversial and have had mixed reviews in the recent literature. Immunosuppressed patients and those receiving chemotherapy are particularly at risk for infectious complications and are therefore given high priority to receiving prophylactic vaccines. METHOD: We administered the influenza vaccine to 29 patients with malignant lymphoma who were receiving chemotherapy or had recently completed therapy during the flu season of 2003-2004. An aged-matched control group received the same vaccine during the same period. The ability of both groups to mount a protective titer of antibodies to the antigens in the vaccine was measured. RESULTS: Three of 29 patients (10%) in the lymphoma group were able to mount a 4-fold titer to at least one of the influenza A antigens. One patient developed a protective titer to both influenza A and B antigens and 3 of 29 responded to the influenza B antigen. In the control group 13 of 29 (45%) responded to an influenza A antigen and 14 of 29 (48%) had a 4-fold response to the B antigen. Seven of 29 controls (24%) had a 4-fold increase in their titers to both the A and B antigens. CONCLUSIONS: This study confirmed the low incidence of response or efficacy to the influenza vaccine reported in previous studies. Only a small percentage (10%) of immunosuppressed patients with malignant lymphoma responded with a 4-fold increase in their antibody titer to the major antigens of the 2003 influenza vaccine. Most interestingly, less than 50% of the aged-matched control population studied responded with a 4-fold increase in their antibody titer. Additional studies are needed to determine methods for improving the efficacy of the vaccine and the effectiveness of the influenza vaccination program in preventing influenza infections in the United States.

Adult↗

The detection of monkeypox in humans in the Western Hemisphere.

BACKGROUND: During May and June 2003, an outbreak of febrile illness with vesiculopustular eruptions occurred among persons in the midwestern United States who had had contact with ill pet prairie dogs obtained through a common distributor. Zoonotic transmission of a bacterial or viral pathogen was suspected. METHODS: We reviewed medical records, conducted interviews and examinations, and collected blood and tissue samples for analysis from 11 patients and one prairie dog. Histopathological and electron-microscopical examinations, microbiologic cultures, and molecular assays were performed to identify the etiologic agent. RESULTS: The initial Wisconsin cases evaluated in this outbreak occurred in five males and six females ranging in age from 3 to 43 years. All patients reported having direct contact with ill prairie dogs before experiencing a febrile illness with skin eruptions. We found immunohistochemical or ultrastructural evidence of poxvirus infection in skin-lesion tissue from four patients. Monkeypox virus was recovered in cell cultures of seven samples from patients and from the prairie dog. The virus was identified by detection of monkeypox-specific DNA sequences in tissues or isolates from six patients and the prairie dog. Epidemiologic investigation suggested that the prairie dogs had been exposed to at least one species of rodent recently imported into the United States from West Africa. CONCLUSIONS: Our investigation documents the isolation and identification of monkeypox virus from humans in the Western Hemisphere. Infection of humans was associated with direct contact with ill prairie dogs that were being kept or sold as pets.

Adolescent↗

The identification of genetically related bacterial isolates using pulsed field gel electrophoresis on nursing home units: a clinical experience.

OBJECTIVES: To describe a laboratory-based technique to track nursing home infections. DESIGN: Retrospective data analysis. SETTING: A 721-bed skilled care facility with 14 nursing units. PARTICIPANTS: Residents in a nursing home, average age 76+/-10, 78% male. MEASUREMENTS: Bacterial isolates were listed for each nursing unit. Clusters of identical species and antibiotic susceptibility were identified followed by pulsed-field gel electrophoresis (PFGE). If the genetic analysis yielded related strains, the director of nursing performed a clinical investigation. PFGE is available through reference laboratories at a cost of approximately 75 dollars/isolate. RESULTS: Twenty-four clinical clusters of phenotypically identical bacteria (species, antibiotic susceptibility) were identified. Fourteen included genetically related isolates. CONCLUSION: Approximately half of the phenotypically identical clusters contained genetically related isolates. The identification of genetically related bacterial isolates on nursing units by PFGE provides staff with a specific circumstance to review secretion precautions. Genetic analysis may also demonstrate that apparent clusters are unrelated.

Aged↗

Novel polymorphisms in mec genes and a new mec complex type in methicillin-resistant Staphylococcus aureus isolates obtained in rural Wisconsin.

We determined allelic polymorphisms in the mec complexes of 524 methicillin-resistant Staphylococcus aureus isolates by partial or complete sequencing of three mec genes, mecA, mecI, and mecR1. The isolates had been collected over a 10-year period from patients living in rural Wisconsin, where the use of antibiotics was expected to be lower than in the bigger cities. Of the 18 mutation types identified, 16 had not been described previously. The five most common mutations were a mutation 7 nucleotides (nt) upstream from the start site (G-->T) in the mecA promoter (34.7%), an E246G change encoded by mecA (2.2%), a cytosine insertion at codon 257 in mecA (13.2%), an N121K change encoded by mecI (7.8%), and a major mecI-mecR1 deletion designated as a class B1 mec complex deletion type (25.4%). There was a high degree of conservation of the amino acid sequence of MecR1. Strains with the same mutations had variable resistance to oxacillin, and the median MIC for isolates that harbored the 7-nt-upstream mutation was lower than that for strains harboring other mutations. Our data suggest that the mecA promoter mutation plays a more important role in determining methicillin resistance than mutations in mecI and mecA do. Eighty-five percent of the tested isolates (n = 148) with the mecA promoter mutation and the class B1 mec complex deletion belonged to the same major clonal group, identified as MCG-2, and harbored the type IV staphylococcal cassette chromosome mec DNA. There was also a wide range of oxacillin MICs for strains with wild-type mecA, mecI, and mecR1 sequences, suggesting that the genetic backgrounds of clinical strains are significant in determining susceptibility to methicillin.

Alleles↗

Isolation of a fastidious Bergeyella species associated with cellulitis after a cat bite and a phylogenetic comparison with Bergeyella zoohelcum strains.

Bergeyella zoohelcum is an uncommon zoonotic pathogen typically associated with cat or dog bites. Previously, only five cases of B. zoohelcum infection have been reported. We report the isolation and characterization of a fastidious Bergeyella species from acute cellulitis in the upper extremity of a 60-year-old woman. The organism was too fastidious for identification and susceptibility testing with traditional culture methods. The isolate was characterized further by PCR amplification and sequencing of the 16S rRNA gene with broad-range eubacterial primers. Phylogenetic analysis of the 16S ribosomal DNA sequence indicated that this isolate was a member of the species B. zoohelcum (previously Weeksella zoohelcum), a gram-negative bacillus that is rarely associated with infections in humans. Despite sharing a close genetic relationship with other B. zoohelcum strains, this isolate was extremely fastidious in nature, raising the possibility that similar strains from cat or dog bite wound infections have been underreported.

Acute Disease↗

Emergence and spread of community-associated methicillin-resistant Staphylococcus aureus in rural Wisconsin, 1989 to 1999.

We investigated the emergence and spread of community-associated strains of methicillin-resistant Staphylococcus aureus (CA-MRSA) in central and northern Wisconsin by determining the temporal and clonal relationships and geographic expansion among 581 of 956 clinical isolates of MRSA collected between 1989 and 1999. Based on EcoRI plasmid profiles (PP), two types, PP-11 and PP-13, were highly stable over time and were consistently associated with multidrug-sensitive strains recovered from outpatients treated at Native American community clinics. Pulsed-field gel electrophoresis (PFGE) yielded six major clonal groups (MCGs) and 19 minor clonal groups. The six MCGs represented 82% of the isolates. All strains with either PP-11 or -13 were present in MCG-2. Eighty-nine percent of the isolates in MCG-2 originated from Native American clinics, and 90% belonged to two PFGE types (19 and 20), the types associated with an outbreak of MRSA in a Native American community in 1992. MCG-2 isolates were multidrug sensitive, harbored type IVa staphylococcal cassette chromosome mec, and were very closely related by PFGE to the Midwestern CA-MRSA strain MW2. MCG-2 strains were mostly obtained from skin infections and affected patients with a mean age of 24 (+/-18.0) years. MCG-2 strains spread to four additional Native American communities and 20 other communities. Our findings suggest that CA-MRSA in Wisconsin likely originated in Native American communities in the early 1990s and since has become widespread throughout the state. Two early CA-MRSA strains (WI-33 and WI-34) in Wisconsin represent progenitors of the MW2 strain, based on their almost indistinguishable genotypic characteristics.

Community-Acquired Infections↗

Molecular characteristics of nosocomial and Native American community-associated methicillin-resistant Staphylococcus aureus clones from rural Wisconsin.

In central and northern Wisconsin methicillin-resistant Staphylococcus aureus (MRSA) was first detected in 1989. Over the next 10-year period, 581 MRSA isolates were collected, 17.2% of which came from patients who were treated at five Native American clinics. These isolates were typed by SmaI-macrorestricted pulsed-field gel electrophoresis (PFGE). The PFGE patterns clustered the isolates into six major clonal groups (MCGs), i.e., MCGs 1 to 6, and 19 minor clonal groups (mCGs). The 25 clonal groups were represented by 109 unique PFGE types. Sixty-five percent of the MCG-2 isolates were recovered from patients who were treated at Native American clinics. Ninety-four percent of the MCG-2 isolates harbored the staphylococcal cassette chromosome mec (SCCmec) IVa. These isolates also had PFGE profiles that were clonally related to the midwestern community-associated MRSA (CA-MRSA) strain, MW2. The representative isolates from MCG-2 had the multilocus sequence type allelic profile 1-1-1-1-1-1-1 and contained pvl genes. They were also susceptible to various antibiotics, a finding consistent with the CA-MRSA phenotype. SCCmec IV was also present in other mCGs. Unlike MCG-2, isolates from the remaining five MCGs harbored SCCmec II and were resistant to multiple antibiotics, suggesting their nosocomial origin. The 19 mCGs were represented by diverse SCCmec types and three putative new variants referred to as SCCmec Ib, IIa, and IIb.

Community-Acquired Infections↗