Detecting Blastocystis using parasitologic and DNA-based methods: a comparative study.

Few studies have targeted the relative performance of diagnostic methods used for the detection of Blastocystis, a unicellular organism often present in fecal specimens from individuals with and without gastrointestinal symptoms. Aims of this study included a comparison of the formol ethyl acetate concentration technique (FECT), permanent trichrome staining of feces fixed in sodium acetate-acetic acid-formalin (SAF-PST), xenic in vitro culture (XIVC), and polymerase chain reaction (PCR) regarding Blastocystis screening of 107 samples from 93 patients with suspected enteroparasitic disease. Compared with PCR, the sensitivity/specificity of XIVC, SAF-PST, and FECT was 89%/100%, 82%/100%, and 50%/100%, respectively.

Multicentre determination of quality control strains and quality control ranges for antifungal susceptibility testing of yeasts and filamentous fungi using the methods of the Antifungal Susceptibility Testing Subcommittee of the European Committee on Antimicrobial Susceptibility Testing (AFST-EUCAST).

A multicentre study involving seven laboratories was performed using techniques recommended by the Antifungal Susceptibility Testing Subcommittee of the European Committee on Antimicrobial Susceptibility Testing (AFST-EUCAST) to evaluate and propose quality control ranges and strains for susceptibility testing of fermentative yeasts and filamentous fungi. Participating laboratories tested the susceptibilities of a panel of 12 encoded isolates to amphotericin B, flucytosine, fluconazole, itraconazole, voriconazole and posaconazole. In total, 15 lots of assay medium were tested, with one lot being common to all laboratories, and 18 144 MIC values were determined.

The prevalence of Dientamoeba fragilis in patients with suspected enteroparasitic disease in a metropolitan area in Denmark.

The prevalence of Dientamoeba fragilis in patients from a metropolitan area in Denmark was determined by examination of paired stool samples using two techniques: a formol ethyl-acetate concentration technique with unpreserved faeces and a permanent staining technique on faeces preserved with sodium acetate-acetic acid-formalin (SAF). Using the SAF permanent staining technique and the formol ethyl-acetate concentration technique, 25% and 15% of the specimens, respectively, were parasite-positive. D.

In vivo efficacy and pharmacokinetics of voriconazole in an animal model of dermatophytosis.

The standard treatment for tinea capitis caused by Microsporum species for many years has been oral griseofulvin, which is no longer universally marketed. Voriconazole has been demonstrated to inhibit growth of Microsporum canis in vitro. We evaluated the efficacy and tissue pharmacokinetics of oral voriconazole in a guinea pig model of dermatophytosis.

Diagnostics of fungal infections in the Nordic countries: we still need to improve!

A Nordic External Quality Assessment programme in medical mycology was established in 2005. In order to monitor not 'best practice' but the level of routine diagnostics, specimens were designed to resemble clinical samples and laboratories were asked to handle the samples like routine samples. Five simulated clinical samples were distributed to 59 participating Nordic laboratories of clinical microbiology.

Five-hour diagnosis of dermatophyte nail infections with specific detection of Trichophyton rubrum.

A rapid two-step DNA extraction method and a multiplex PCR for the detection of dermatophytes in general and Trichophyton rubrum specifically were developed and evaluated with DNA extracted from pure cultures and from clinically diseased nails. DNA from the following dermatophytes was used: Epidermophyton floccosum, Microsporum audouinii, Microsporum canis, Microsporum gypseum, Microsporum nanum, Trichophyton mentagrophytes, Trichophyton rubrum, Trichophyton schoenleinii, Trichophyton soudanense, Trichophyton terrestre, Trichophyton tonsurans, Trichophyton verrucosum, and Trichophyton violaceum. Human DNA and DNA from the following nondermatophyte fungi were included as controls: Alternaria, Aspergillus niger, Candida albicans, Candida glabrata, Candida krusei, Malassezia furfur, Saccharomyces cerevisiae, and Scopulariopsis brevicaulis.

Detection of Blastocystis hominis in unpreserved stool specimens by using polymerase chain reaction.

Blastocystis hominis is a common enteric parasite of worldwide distribution. Its pathogenetic potential has not yet been established, although numerous case reports suggest that B. hominis may cause the development of various gastrointestinal symptoms and disorders.

Acute pulmonary aspergillosis in immunocompetent subjects after exposure to bark chippings.

We describe 2 cases of immunocompetent males with acute community-acquired invasive pulmonary aspergillosis developing shortly after spreading bark chippings. One patient with a fatal outcome was initially diagnosed as allergic alveolitis rather than infection and received steroid treatment illustrating the obvious challenges in the differential diagnosis between these disease entities.

Prevalence of toe nail onychomycosis in diabetic patients.

Onychomycosis among diabetic patients has been reported in some studies to be of high prevalence. This study aimed to investigate the prevalence of onychomycosis among diabetic patients at a Danish University Hospital. Clinical and mycological examinations were performed on type 1 and 2 diabetic patients from in- and out-patient clinics.

Amphotericin B and caspofungin resistance in Candida glabrata isolates recovered from a critically ill patient.

Background: Consecutive Candida glabrata isolates recovered from a patient in an intensive care unit were resistant to amphotericin B (minimum inhibitory concentration, up to 32 mu g/mL; determined by Etest [AB Biodisk]). Analyses at the national reference laboratory showed that some isolates were also resistant to azoles and caspofungin. In this study, 4 isolates were studied thoroughly using susceptibility assays and a mouse model and to determine clonality.

Four cases of Candida albicans infections with isolates developing pink colonies on CHROMagar Candida plates.

Candida albicans, the most commonly isolated yeast species, is typically identified by its green colony-colour on CHROMagar Candida plates. We here report four cases of Candida albicans infections, in which the initial identification was non-albicans isolates due to a clear pink colour of the colonies on CHROMagar Candida plates. However, classical phenotypic criteria, biochemical assimilation pattern and molecular characterisation identified all four isolates as C.

Seminational surveillance of fungemia in Denmark: notably high rates of fungemia and numbers of isolates with reduced azole susceptibility.

The aim of this study was to present the first set of comprehensive data on fungemia in Denmark including the distribution of species and range of susceptibility to major antifungal compounds based on a seminational surveillance study initiated in 2003. The catchment area of the participating hospitals had a population of 2.8 million, or 53% of the Danish population.

Disseminated Penicillium marneffei sepsis in a HIV-positive Thai woman in Denmark.

We report the first case of disseminated Penicillium marneffei infection, in a 32-y-old HIV positive Thai woman, in Denmark. Untreated it is a life-threatening infection. Therefore it is extremely important to consider P.

Fusarium fungaemia in immunocompromised patients.

Fusarium spp. cause infections only rarely in immunologically competent hosts, but disseminated infection may occur in severely immunocompromised patients. Symptoms of disseminated infection are persistent fever, despite broad-spectrum antibacterial and antifungal treatment, associated with skin lesions, most commonly on the extremities, in 60-80% of patients.

Cryptococcus neoformans var neoformans resistant to fluconazole in an HIV-negative patient with chronic lymphocytic leukemia.

Cases of fluconazole-resistant Cryptococcus neoformans have been reported in AIDS patients previously treated with fluconazole. We report a case of fluconazole-resistant cryptococcal meningitis in an HIV-negative patient not previously exposed to fluconazole. The patient experienced a clinical relapse after discontinuation of therapy with amphotericin B and subsequent initiation of fluconazole therapy.

In vivo pathogenicity of eight medically relevant Candida species in an animal model.

Background: The relative pathogenicity of eight medically important Candida species was investigated in a mouse model.

Materials And Methods: Seventeen isolates were included; two isolates of Candida albicans, Candida glabrata, Candida tropicalis, Candida lusitaniae, Candida parapsilosis, Candida kefyr and Candida guilliermondii and three Candida krusei isolates. Mice in groups of three or four were inoculated with 10(5) and 10(7) CFU, respectively.

Prevalence of and detection of resistance to ampicillin and other beta-lactam antibiotics in Haemophilus influenzae in Denmark.

The susceptibility of Haemophilus influenzae to penicillin V and G, ampicillin and cefuroxime was investigated by MIC, disc and tablet diffusion methods, using chocolate agar as test medium, to determine the prevalence of ampicillin-resistant isolates and the optimal method for their detection. Eighty-six isolates were clinical isolates collected prospectively from July to September 1998 and 22 isolates were clinical isolates with decreased susceptibility to ampicillin previously referred to the reference laboratory. Eighty-seven isolates were ampicillin-susceptible and 16 were ampicillin-resistant.

Comparison of Etest and a tablet diffusion test with the NCCLS broth microdilution method for fluconazole and amphotericin B susceptibility testing of Candida isolates.

Three methods were compared for the susceptibility testing of yeast isolates to fluconazole and amphotericin B: two fagar diffusion methods (Etest and a tablet diffusion test) and the National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution method. Given as MIC(50)s (range), fluconazole endpoints were: for the 24 h broth microdilution test, 0.25 mg/L (0.

[Deep wound infection with Salmonella enteritidis following osteosynthesis].

A 43-year old man with alcohol abuse had septicaemia with Salmonella enteritidis. After osteosynthesis of an intertrochanteric femoral fracture he developed a deep wound infection with Salmonella enteritidis. The patient was treated with surgical debridement and antibiotics.

[Spondylitis and medullary transverse syndrome caused by Staphylococcus aureus].

Three cases of haematogenous osteomyelitis in the vertebral column caused by Staphylococcus aureus are reported. The cases, which were associated with severe neurological symptoms and/or death, were initially characterized by a long period with no or discrete local signs of infection and by values of temperature and leucocyte counts within or close to normal values. In this period measurements of the sedimentation reaction and C-reactive protein were elevated, and were markers of persistent infection.

Diagnosing bacteremia at a Danish hospital using one early large blood volume for culture.

The objective of the study was to evaluate the yield of a blood culture based on inoculating 40 ml of blood drawn from 1 venipuncture. During a 3-year period (1990-1992) 1351 (19.3%) of 6994 blood culture sets were positive.

The HIV-1 V3 domain on field isolates: participation in generation of escape virus in vivo and accessibility to neutralizing antibodies.

The V3 domain is highly variable and induces HIV neutralizing antibodies (NA). Here we addressed the issues of 1) the participation of mutations in V3 in generation of neutralization resistant escape virus in vivo and 2) the applicability of synthetic V3 peptides corresponding to field isolates to induce neutralizing immune sera. Seven peptides corresponding to the V3 region of primary and escape virus from 3 HIV-1 infected patients were synthesized and used for antibody (Abs) studies and immunizations.

Neutralization epitopes on HIV pseudotyped with HTLV-I: conservation of carbohydrate epitopes.

One mechanism for expanding the cellular tropism of human immunodeficiency virus (HIV) in vitro is through formation of phenotypically mixed particles (pseudotypes) with human T lymphotropic virus type I (HTLV-I). In this study we found that pseudotypes allow penetration of HIV particles into CD4-negative cells, previously nonsusceptible to HIV infection. The infection of CD4-negative cells with pseudotypes could be blocked with anti-HTLV-I serum but failed to be significantly inhibited with anti-HIV serum or a V3-neutralizing anti-gp120 monoclonal antibody.

HIV-1 infection of in vitro cultured human monocytes: early events and influence of anti HIV-1 antibodies.

To characterize the role of the humoral immune response on HIV-1 infection of monocytes and macrophages (M phi s) we examined the susceptibility of in vitro cultured monocyte/M phi s to various HIV-1 isolates and the influence of heterologous and particularly autologous anti HIV-1 sera on this infection. Depending on the period of in vitro cultivation and the virus isolate used different patterns of susceptibility were detected. One week old monocyte/M phi s were highly susceptible to HIV-1 infection, in contrast to monocyte/M phi s cultured 4 weeks.