Population-based surveillance and a case-control study of risk factors for endemic lymphocutaneous sporotrichosis in Peru.

Population-based surveillance and a case-control study were conducted in Abancay, Peru, to estimate the burden of disease and to determine risk factors for sporadic lymphocutaneous sporotrichosis (LS). Laboratory records from local hospitals were reviewed for the years of 1997 and 1998, and prospective surveillance was conducted for the period of September 1998 through September 1999. A case-control study was conducted with 2 matched control subjects per case patient. The mean annual incidence was 98 cases per 100,000 persons. Children had an incidence 3 times higher than that for adults and were more likely to have LS lesions on the face and neck. Identified risk factors included owning a cat, playing in crop fields, having a dirt floor in the house, working mainly outdoors, and having a ceiling made of raw wood or conditions associated with a lower socioeconomic status. Decreased environmental exposure, such wearing protective clothing during construction activities for adults or limiting contact with cats and soil for children, and improvements in living spaces may decrease the incidence of LS.

Antigen and antibody attachment in ELISA for Aspergillus fumigatus IgG antibodies.

Four commercial antigen extracts of Aspergillus fumigatus were evaluated for use in a rapid enzyme-linked immunosorbent assay (ELISA) for anti-A. fumigatus IgG. Initial binding of both somatic and culture filtrate preparations to a polyvinyl chloride solid phase was concentration dependent and increased with incubation time. Antigen binding to the solid phase was reproducible. Binding of A. fumigatus precipitin-positive serum to bound antigen was rapid. All four A. fumigatus antigens demonstrated similar dose-response curves when tested against pooled sera containing a high titre of A. fumigatus antibodies. Detectable activity in precipitin test-negative sera decreased rapidly with dilution. All the antigen preparations were found to be suitable for ELISA procedures and permit the rapid determination of IgG antibodies to A. fumigatus.

Computer-assisted rapid enzyme-linked immunosorbent assay (ELISA) in the serological diagnosis of aspergillosis.

A computer-assisted method for the positive/negative discrimination of ELISA data is described. This method was applied to a rapid ELISA procedure for IgG class antibodies to Aspergillus fumigatus in which the performance time of the test was reduced to 1 h. The method gives results which compare well with those by agar gel double diffusion (AGDD). The computer-assisted reading, calculation and tabulation of ELISA results from one microtitre plate is performed in less than 2 min and permits analysis of large numbers of specimens.

Clinical pharmacokinetics of ketoconazole.

Ketoconazole, a synthetic imidazole antifungal, is effective for superficial fungal infections, genital candidosis and chronic mucocutaneous candidosis, and has been used in immunocompromised patients and advanced prostatic carcinoma. Absorption of ketoconazole is variable after oral administration, with large variability in peak serum concentrations. Antacids reduce, and food or dilute hydrochloric acid increase, absorption. Renal failure and bone marrow transplantation are associated with reduced absorption. Ketoconazole is not absorbed systemically after topical administration, and minimally absorbed from the vagina. Distribution of ketoconazole varies according to the tissue sampled, the underlying disease and the dose and duration of treatment. Ketoconazole does not cross the intact blood-brain barrier, and crosses to only a limited extent in fungal meningitis. Urinary concentrations of ketoconazole are usually low, but vaginal and vaginal tissue concentrations correlate with those in serum. Seminal fluid concentrations are inadequate for treatment of epididymitis. Ketoconazole is 83.7% plasma protein (mainly albumin) bound, and 15.3% is erythrocyte bound, resulting in only 1% of free drug. Animal studies indicate strong binding to the cytochrome P-450 mono-oxygenase complex. Extensive metabolism to inactive metabolites occurs, the products being mainly excreted in the faeces. Saturable hepatic first-pass metabolism is probable. The half-life of ketoconazole is dose-dependent, increases during long term treatment, suggesting auto-inhibition of metabolism. The kinetics after oral administration fit a 2-compartment model. Drug interactions of theoretical, if not practical, significance include warfarin, chlordiazepoxide, methylprednisolone, cyclosporin and drugs known to induce microsomal enzymes. In each case, some dosage adjustment for ketoconazole, or the interacting drug, may be required.

Indirect immunofluorescence test for the detection of Aspergillus fumigatus antibodies.

An indirect immunofluorescence test was both sensitive and specific in detecting IgG antibodies to Aspergillus fumigatus. The test revealed a significant increase in titre of antibodies in patients with the allergic or mycetomal forms of pulmonary aspergillosis.

Detection of Aspergillus fumigatus precipitins: a comparison of counter immunoelectrophoresis and double diffusion.

A counter immunoelectrophoresis method was compared with a double diffusion test for detecting Aspergillus fumigatus antibodies. Of 70 patients, 23 gave positive results in both tests and the remainder gave negative results. Fifteen patients with proven aspergillosis gave positive results in both tests.

Observations on the use of the double diffusion test in the diagnosis of vaginal candidiasis.

Precipitins to mannan and cytoplasmic antigens of three Candida species were determined in an unselected series of 289 non-pregnant women. Precipitins were present in 20% of sera of women with vaginal candidiasis, in 23% of women harbouring yeasts in the vagina without clinical signs of infection, and in 21% of women harbouring yeasts in sites other than the vagina. Of the 47 patients who reacted with Candida albicans mannan or cytoplasmic antigens, 98% reacted with mannan antigen but only 13% with cytoplasmic antigen. The inclusion of mannan and cytoplasmic antigens of C. guilliermondii and C. parapsilosis did not increase the specificity or sensitivity of the test in the diagnosis of vaginal candidiasis. It is suggested that the double diffusion test is of doubtful value as an adjunct to the diagnosis of vaginal candidiasis.

Rapid enzyme-linked immunosorbent assay (ELISA) for Aspergillus fumigatus antibodies.

A rapid enzyme-linked immunosorbent assay (ELISA) where component incubation periods were shortened to one hour, was compared with agar gel double diffusion (AGDD) and a standard ELISA procedure for detecting antibodies to Aspergillus fumigatus in 28 asthmatic patients with suspected allergic aspergillosis. Using two A fumigatus antigens the rapid ELISA compared well with AGDD and the standard ELISA method. Eleven sera that reacted with both antigens in AGDD were all positive against antigen 1 in both forms of ELISA, but two failed to react with antigen 2 in the standard ELISA and three failed to react with this antigen in the rapid method. Thirteen AGDD-negative sera were also negative in both forms of ELISA. The rapid ELISA provides a sensitive and reproducible test for routine serological investigation of allergic aspergillosis.

Serological diagnosis of infection of the urinary tract by yeasts.

Precipitins to mannan and cytoplasmic antigens of Candida albicans and Torulopsis glabrata were determined in 25 patients with colonization of the bladder urine by yeasts and in 25 control patients with bacteriuria. Precipitins were present in 64% of the patients with funguria and in 56% of the patients with bacteriuria. There was no correlation between the reactions obtained and the clinical significance of the yeast colonization. It is suggested that a single precipitin test is unhelpful in the assessment of the significance of urinary colonization by yeasts in hospital patients.

Evaluation of the AUXACOLOR system, a new method of clinical yeast identification.

AIMS: To compare the AUXACOLOR yeast identification system with the API 20C system. METHODS: Yeast isolates (n = 215), comprising 16 species, were identified using the AUXACOLOR system and the API 20C system. Isolates that could not be identified with the API 20C system or which produced discrepant results in the two systems were identified by assimilation and fermentation procedures. RESULTS: AUXACOLOR correctly identified 150 (85.7%) of 175 germ tube negative isolates while API 20C identified 155 (88.6%). Incorrect identifications were more common with API 20C (7.4%) than with AUXACOLOR (3.7%). Of 110 isolates of four common pathogens (Candida glabrata, C parapsilosis, C tropicalis, and Cryptococcus neoformans), 82.7% (91/110) were identified by AUXACOLOR while API 20C identified 74.5% (82/110). Of 65 less common germ tube negative isolates, 55.4% (36/65) were identified by AUXACOLOR while API 20C identified 63.1% (41/65). CONCLUSION: Although it has a limited database of 26 species, the AUXACOLOR system is a useful method for identification of germ tube negative clinical yeast isolates. Compared with the API 20C, the AUXACOLOR system is simpler and quicker to set up, easier to interpret, and comparable in cost.

Aspergillus antigen testing in bone marrow transplant recipients.

AIMS: To assess the clinical usefulness of a commercial aspergillus antigen enzyme linked immunosorbent assay (ELISA) in the diagnosis of invasive aspergillosis (IA) in bone marrow transplant recipients, and to compare it with a commercial latex agglutination (LA) test. METHODS: In total, 2026 serum samples from 104 bone marrow transplant recipients were tested. These comprised 67 sera from seven patients who had died with confirmed IA, 268 sera from nine patients who had died with suspected IA, and 1691 sera from 88 patients with no clinical, radiological, or microbiological signs of IA. RESULTS: The ELISA was more sensitive than the LA test. All patients who were ELISA positive were also LA positive, and a positive LA result never preceded a positive ELISA. Twelve of 16 patients with confirmed or suspected IA were ELISA positive on two or more occasions, compared with 10 of 15 who were LA positive. ELISA was positive before LA in five patients (range, 2-14 days), and became positive on the same day in the remainder. Aspergillus antigen was detected by ELISA a median of 15 days before death (range, 4-233). Clinical and/or radiological evidence of IA was noted in all patients, and a positive ELISA was never the sole criterion for introduction of antifungal treatment. Two samples (one from each of two patients without IA) gave false positive results. CONCLUSIONS: The aspergillus ELISA is a specific indicator of invasive aspergillosis if the criterion of two positive samples is required to confirm the diagnosis. However, the test is insufficiently sensitive to diagnose aspergillosis before other symptoms or signs are apparent, and hence is unlikely to lead to earlier initiation of antifungal treatment. It is therefore unsuitable for screening of asymptomatic patients at risk of invasive aspergillosis, but does have a useful role in confirming the diagnosis in symptomatic patients.

Use of the Pastorex aspergillus antigen latex agglutination test for the diagnosis of invasive aspergillosis.

AIMS: To evaluate the Pastorex aspergillus antigen latex agglutination test for the diagnosis of invasive aspergillosis in patients undergoing liver or bone marrow transplantation. METHODS: Serum samples were taken at least twice weekly post-transplant and tested for Aspergillus antigen. Latex agglutination test results were compared with microbiological examination of respiratory, urine and bile specimens. Serum samples from liver transplant patients were also tested for antibodies to Aspergillus fumigatus by counter immunoelectrophoresis. RESULTS: Eight of the 91 patients studied developed invasive aspergillosis. Positive latex agglutination tests were obtained in eight of 187 (4.3%) serum samples from four of these eight patients. The other four patients with invasive aspergillosis gave consistently negative latex agglutination tests. A positive latex agglutination test was the first indication of invasive aspergillosis in two patients; these patients were already on amphotericin B. Positive latex agglutination tests were the only evidence of invasive aspergillosis in one patient who subsequently died of the infection. False positive latex agglutination tests were obtained in five of 83 (6%) patients with no evidence of invasive aspergillosis and misleading positive cultures seen in nine of 83 (10.8%). No antibodies were detected in three of four liver transplant patients with invasive aspergillosis. Conversely, antibodies were detected in 63 of 262 (24%) serum samples from 43 liver transplant patients with no evidence of invasive aspergillosis; one of these patients had an antibody titre of 1:2 on four separate occasions. CONCLUSIONS: The Pastorex aspergillus antigen latex agglutination test, when used alone, lacks sensitivity and specificity for the early diagnosis of invasive aspergillosis. A diagnosis was made in all patients with invasive aspergillosis when both culture and antigen tests were performed although using these criteria a false positive diagnosis would have been made in 13 of 83 (15.6%) patients. Microbiological and serial serological investigations for antigen should both be performed and the results considered in conjunction with radiological and clinical evidence.

Chaetomium pneumonia in patient with acute myeloid leukaemia.

A patient with relapsed refractory acute myeloid leukaemia developed typical fungal lung lesions despite intravenous amphotericin B prophylaxis. Chaetomium globosum was cultured from the resected right lower lobe. Histology showed branching hyphae negative for common Aspergillus species by immunohistochemical staining. Previous reports of invasive disease caused by Chaetomium and some applications of immunohistochemical staining for Aspergillus are discussed.

Treatment of vaginal candidiasis with miconazole.

An open trial was carried out in 50 women with clinical signs and mycological evidence of vaginal candidiasis to compare the effectiveness of treatment with 2% miconazole cream with that of nystatin pessaries. Both groups were given intravaginal treatment for 14 days and were followed up at monthly intervals for 3 months. Relief of symptoms and clinical signs was comparable in the two groups but the mycological cure rate at 1 month was higher with miconazole. At 2 and 3 months, however, there was no significant difference between the cure rate obtained with the two treatments in the patients still included in the study.

Rhizopus microsporus in wooden tongue depressors: a major threat or minor inconvenience?

The investigation and management of an apparent outbreak of Rhizopus spp. in a London paediatric referral centre between September 1995 and April 1996 is described. The organism was identified in microbiological surveillance samples from 23 patients nursed in four hospital areas. Investigations revealed the presence of the organism in spatulae from all ward areas investigated and from closed boxed containers held in the central hospital stores obtained from a new supplier. In contrast, culture of spatulae from the initial supplier failed to yield any fungal isolates. The incident was reported to the Medical Device Agency (MDA), the Central Public Health Laboratory Service (CPHLS) and the Birmingham PHLS. A statement was prepared for the weekly Communicable Disease Report and a hazard warning issued by the MDA. The spatulae were withdrawn from use and the contract with the original supplier was re-established. This incident resulted in contamination of samples only and no patient involvement. It highlights the problems which may follow use of equipment for unintended purposes and the need for good manufacturing practice guidelines to be applied to non-sterile equipment used in direct patient care.

An outbreak of aspergillosis in a general ITU.

Over a 5-month period, six patients in a general intensive therapy unit became colonized by Aspergillus species including Aspergillus fumigatus, and invasive infection occurred in at least two of them. Broncho-alveolar lavage was unhelpful in discriminating between infection and colonization. The source of infection was presumed to be disturbance of an accumulation of spores in fibrous insulation material above the perforated metal ceiling. Patients in such units without clearly identifiable defects of defence against infection may be at risk from aspergillosis. The risk can be reduced by improved hospital design, satisfactory ventilation and thorough regular cleaning of environmental surfaces.

Treatment failure in a case of fungal keratitis caused by Pseudallescheria boydii.

A case is presented of Pseudallescheria boydii fungal keratitis in an agricultural welder. Treatment with azole antifungal drugs (miconazole and itraconazole) and with penetrating keratoplasty was unsuccessful in eradicating the infection, and eventually the eye was eviscerated.

Acremonium infection in a neutropenic patient.

Acremonium species (previously Cephalosporium) are saprophytic moulds of low pathogenicity commonly found in the environment. We report a case of repeated acremonium fungaemia in a neutropenic patient, probably associated with a Hickman line. Fungaemia was accompanied by fever and a papular skin rash, both of which appeared to resolve when the neutrophil count recovered without antifungal chemotherapy, only to return when the patient next became neutropenic.

Fatal Candida tropicalis fungaemia in a leukaemic patient receiving fluconazole prophylaxis.

A 50-year-old man with newly diagnosed acute myeloid leukaemia developed breakthrough candidaemia while receiving fluconazole as antifungal prophylaxis during remission-inducing chemotherapy. Candida tropicalis was isolated; the strain was resistant to fluconazole on in vitro sensitivity testing, a phenomenon which has not been previously reported in this setting.

Disseminated infection due to Cylindrocarpon lichenicola in a patient with acute myeloid leukaemia.

We describe what is to our knowledge the first reported case of disseminated infection due to Cylindrocarpon. The presumed source was athlete's foot, a condition with which this fungus has previously been associated. Diagnosis was made by needle aspiration of a cutaneous lesion. Radiographic evidence of pulmonary involvement was present. The infection resolved following marrow regeneration and treatment with amphotericin B. Correct identification of Cylindrocarpon may be useful in guiding antifungal therapy.