R. Kappe

Disseminated Penicillium marneffei infection in an HIV-positive female from Thailand in Germany.

We report the case of a 33 year old Thai female, who was married in Germany for eight years and used to travel to Thailand every year for several weeks. She presented with abdominal and back pain, prolonged fever, generalized lymphadenopathy, and a recent history of oral thrush. She was diagnosed HIV positive with initial CD4 counts of 18/μl and an HI virus load of 59.000 copies/ml. Antiviral therapy was installed with zidovudin, lamivudin, and efavirenz. Abdominal CT scans revealed greatly enlarged abdominal lymphnodes. Fine needle aspirates of cervical and retroperitoneal lymphnodes, sputum samples, blood samples, and a bone marrow biopsy were microscopically positive for Penicillium marneffei and grew P. marneffei. The isolates were sensitive to amphotericin B, flucytosine, itraconazole, and fluconazole. Both universal and specific fungal polymerase chain reaction assays were positive in various samples. Serum Aspergillus galactomannan antigen, which is known to crossreact with P. marneffei, was elevated and subsequently used for monitoring of therapy. With antifungal treatment (intravenous amphotericin B 0.6 mg/kg/d for two weeks, oral itraconazole 400 mg/d for 10 weeks and 200 mg/d as maintenance therapy), the fever declined in 6 days, the size of the enlarged lymphnodes gradually decreased in the CT scans, and the initial abdominal and back pain vanished.

Polymerase chain reaction-based detection of dermatophyte DNA with a fungus-specific primer system

Summary. There is significant clinical interest in primers which are specific for fungi and do not hybridize to DNA of other eukaryotes or prokaryotes. Such primers would allow specific amplification of fungal DNA from human tissue samples containing fungi. Fungal identification to the species level could follow by direct sequencing or restriction analysis. Several previously described primer systems cross‐react with DNA of plants and animals.

In vitro susceptibility of 15 strains of zygomycetes to nine antifungal agents as determined by the NCCLS M38-A microdilution method.

The in vitro susceptibility of 15 strains of six genera of zygomycetes including Rhizopus oryzae (Rhizopus arrhizus), Rhizopus stolonifer, Mucor circinelloides (three), Absidia corymbifera (three), Rhizomucor pusillus (three), Cunninghamella bertholletiae (two), and Syncephalastrum racemosum (two) to nine antifungal agents were determined by the NCCLS M38‐A broth microdilution method. Geometric means of the minimal inhibitory concentrations (MIC) were: amphotericin B 0.07 mg l−1 (range 0.03–0.5 mg l−1), nystatin 0.83 mg l−1 (range 0.25–4 mg l−1), itraconazole 0.59 mg l−1 (range 0.03 to >8 mg l−1), voriconazole 6.50 mg l−1 (range 2 to >8 mg l−1), ciclopiroxolamine 1.59 mg l−1 (range 0.5–4 mg l−1), and amorolfine 9.19 mg l−1 (range 1 to >16 mg l−1). All strains were resistant to 5‐flucytosine, fluconazole (MIC >64 mg l−1) and caspofungin (MIC >16 mg l−1).

Performance of an IS6110-Based PCR Assay and the COBAS AMPLICOR MTB PCR System for Detection of Mycobacterium tuberculosis Complex DNA in Human Lymph Node Samples

ABSTRACT We compared the performance of two PCR assays, an IS6110-based in-house protocol and the COBAS AMPLICOR MTB PCR (COBAS MTB) system, for the detection of Mycobacterium tuberculosis complex in 43 human lymph node samples from 40 patients. For the in-house PCR and the COBAS MTB assays, respectively, sensitivities were 87.5% versus 45.5% (P < 0.05), specificities were 100.0% versus 91.3% (P > 0.05), and inhibition rates were 4.8% versus 19.5% (P < 0.05). For the COBAS MTB system, additional N-acetyl-l-cysteine-NaOH pretreatment of the samples changed neither the inhibition rate nor the sensitivity significantly.

Karyotyping of Candida albicans and Candida glabrata from patients with Candida sepsis.

The aim of this study was to determine the relatedness of Candida strains from patients suffering from Candida septicaemia by typing of Candida isolates from blood cultures and different body sites by pulsed field gel electrophoresis (PFGE using a contour‐clamped homogenous electric field, CHEF). We studied 17 isolates of Candida albicans and 10 isolates of Candida glabrata from six patients. Four patients suffered from a C. albicans septicaemia, one patient from a C. glabrata septicaemia, and one patient had a mixed septicaemia with C. albicans and C. glabrata. Eight isolates from blood cultures were compared with 19 isolates of other sites (stool six, urine four, genital swab four, tip of central venous catheter three, tracheal secretion one, sputum one). PFGE typing resulted in 10 different patterns, four with C. albicans and six with C. glabrata. Five of the six patients had strains of identical PFGE patterns in the blood and at other sites. Seven isolates of a 58‐year‐old female with a C. glabrata septicaemia fell into five different PFGE patterns. However, they showed minor differences only, which may be due to chromosomal rearrangements within a single strain. Thus it appears, that the colonizing Candida strains were identical to the circulating strains in the bloodstream in at least five of six patients.

Cross-reactivity of the PLATELIA CANDIDA antigen detection enzyme immunoassay with fungal antigen extracts.

ABSTRACT We studied the specificity of the PLATELIA CANDIDA Ag enzyme immunoassay by using 130 isolates of 63 clinically relevant fungal species. Antigen extracts of seven Candida spp. (Candida albicans, C. dubliniensis, C. famata, C. glabrata, C. guilliermondii, C. lusitaniae, and C. tropicalis) repeatedly yielded positive reactions (>0.5 ng/ml). Geotrichum candidum and Fusarium verticillioides were found to yield borderline-positive reactions (0.25 to 0.50 ng/ml). Antigen preparations from the other 54 fungal species, including yeasts, molds, dermatophytes, and dimorphic fungi, did not cross-react in the assay.

Performance of the Candida mannan antigen detection in patients with fungemia

Zur Diagnostik invasiver Candidosen steht seit einigen Jahren der kommerziell erhältliche Platelia®Candida Mannan Antigen Enzymimmunoassay (Candida EIA) zur Verfügung. Wir evaluierten den Candida EIA an Patienten mit nachgewiesener Fungämie durch Sproßpilze, von denen zumindest eine Serumprobe zur Verfügung stand. In die Studie eingeschlossen wurden 59 Patienten mit 121 Serumproben, von denen 61 verschiedene Sproßpilz‐Isolate aus Blutkulturen angezüchtet wurden. Der Candida EIA war bei 43 von 59 Fungämie‐Patienten positiv (n = 35) oder grenzwertig (n = 8), woraus eine Sensitivität von 73% resultierte. Für die einzelnen Sproßpilz‐Arten ergaben sich folgende Positiv‐Raten: Candida albicans 30 von 39 (77%), Candida glabrata 7 von 11 (64%), Candida parapsilosis 1 von 3, Candida tropicalis 2 von 2, Candida kefyr 2 von 2, Candida lipolytica 0 von 1, Candida lusitaniae 1 von 1, Candida krusei 1 grenzwertig von 1, Saccharomyces cerevisiae 1 von 1. Bei 6 Patienten war der zeitliche Antigen‐Verlauf auswertbar. In 3 Fällen war der Antigennachweis 3 bis 4 Tage vor dem Tag der Blutkulturentnahme positiv, in einem Fall am selben Tag und in 2 Fällen 2 bzw. 5 Tage danach.

Intrinsic in vitro susceptibility of primary clinical isolates of Aspergillus fumigatus, Aspergillus terreus, Aspergillus nidulans, Candida albicans and Candida lusitaniae against amphotericin B

A total of 60 clinical fungal isolates from patients without prior amphotericin B treatment and three control strains were evaluated for their intrinsic susceptibility to amphotericin B (AmB) using microdilution, Etest and disc diffusion assays, on three media each, Roswell Park Memorial Institute (RPMI) 1640, Antibiotic Medium 3 (AM3) and High Resolution Medium. The fungal strains included isolates of Aspergillus fumigatus (n = 10), Aspergillus terreus (n = 12), Aspergillus nidulans (n = 9), Candida albicans (n = 6) and Candida lusitaniae (n = 23). The A. terreus strains were significantly less susceptible to AmB than the A. fumigatus strains in all nine experimental settings (P‐values ranging from 0.009 to <0.00001). The A. nidulans strains were equally susceptible to AmB as the A. fumigatus strains in seven of nine experimental settings and less susceptible in two (microdilution performed on RPMI and AM3, P = 0.01 and 0.007). The C. lusitaniae strains were equally susceptible to AmB as the C. albicans strains in seven of nine experimental settings and more susceptible in two (microdilution and Etest, both performed on AM3, P = 0.01 and 0.0002). Thus, we confirmed that A. terreus is intrinsically less susceptible to AmB than A. fumigatus. In contrast, nine German clinical isolates of Aspergillus nidulans were found equally susceptible to AmB as 10 isolates of A. fumigatus. Furthermore, we found 23 German clinical isolates of C. lusitaniae from patients without prior treatment with AmB equally susceptible to AmB as C. albicans.

Performance des Candida-Mannan-Antigennachweises bei Patienten mit Fungämien

Zur Diagnostik invasiver Candidosen steht seit einigen Jahren der kommerziell erhältliche Platelia®Candida Mannan Antigen Enzymimmunoassay (Candida EIA) zur Verfügung. Wir evaluierten den Candida EIA an Patienten mit nachgewiesener Fungämie durch Sproßpilze, von denen zumindest eine Serumprobe zur Verfügung stand. In die Studie eingeschlossen wurden 59 Patienten mit 121 Serumproben, von denen 61 verschiedene Sproßpilz‐Isolate aus Blutkulturen angezüchtet wurden. Der Candida EIA war bei 43 von 59 Fungämie‐Patienten positiv (n = 35) oder grenzwertig (n = 8), woraus eine Sensitivität von 73% resultierte. Für die einzelnen Sproßpilz‐Arten ergaben sich folgende Positiv‐Raten: Candida albicans 30 von 39 (77%), Candida glabrata 7 von 11 (64%), Candida parapsilosis 1 von 3, Candida tropicalis 2 von 2, Candida kefyr 2 von 2, Candida lipolytica 0 von 1, Candida lusitaniae 1 von 1, Candida krusei 1 grenzwertig von 1, Saccharomyces cerevisiae 1 von 1. Bei 6 Patienten war der zeitliche Antigen‐Verlauf auswertbar. In 3 Fällen war der Antigennachweis 3 bis 4 Tage vor dem Tag der Blutkulturentnahme positiv, in einem Fall am selben Tag und in 2 Fällen 2 bzw. 5 Tage danach.

Fatty acid analysis of different Candida species by capillary column-gas-liquid chromatography

Summary. Gas‐liquid chromatography was used to analyse the fatty acid composition of whole‐cell hydrolysates of 40 yeast strains representing 5 Candida species and Torulopsis glabrata, T. glabrata could be easily distinguished from all other Candida spp. by the absence of C18:3. Candida albicans, C. krusei, C. parapsilosis, C. pseudotropicalis and C. tropicalis showed 10 major peaks in characteristic proportions. C. parapsilosis showed a high C16:0/ C16:l ratio (> 4.5), whereas C. pseudotropicalis was characterized by a C18:1/C16:0 ratio of < 2.0. A high C18:3 concentration (> 10%) was typical for C. krusei (C18:2/C18:3 ratio ≤ 1.0). Our data reveal characteristic patterns of cellular fatty acid composition of T. glabrata, C. krusei, C. parapsilosis and C. pseudotropicalis which can be used for laboratory identification.