Mucormycosis is a rare and opportunistic infection caused by fungi belonging to the order Mucorales.Recent reports have demonstrated an increasing incidence of mucormycosis, which is frequently lethal, especially in patients suffering from severe underlying conditions such as immunodeficiency. In addition, even though conventional mycology and histopathology assays allow for the identification of Mucorales, they often fail in offering a species-specific diagnosis. Due to the lack of other laboratory tests, a precise identification of these molds is thus notoriously difficult. In this study we aimed to develop a molecular biology tool to identify the main Mucorales involved in human pathology. A PCR strategy selectively amplifies genomic DNA from molds belonging to the genera Absidia, Mucor, Rhizopus, and Rhizomucor, excluding human DNA and DNA from other filamentous fungi and yeasts. A subsequent digestion step identified the Mucorales at genus and species level. This technique was validated using both fungal cultures and retrospective analyses of clinical samples. By enabling a rapid and precise identification of Mucorales strains in infected patients, this PCR-restriction fragment length polymorphism-based method should help clinicians to decide on the appropriate treatment, consequently decreasing the mortality of mucormycosis.

The ability of cyclodextrins to enhance the antiviral activity of a phosphodiester oligodeoxynucleotide has been investigated. A 18-mer oligodeoxynucleotide complementary to the initiation region of the mRNA coding for the spike protein and containing the intergenic consensus sequence of an enteric coronavirus has been tested for antiviral action against virus growth in human adenocarcinoma cells. The phosphodiester oligodeoxynucleotide only showed a limited effect on virus growth rate (from 12 to 34% viral inhibition in cells treated with 7.5 to 25 microM oligodeoxynucleotide, respectively, at a multiplicity of infection of 0.1 infectious particle per cell). In the same conditions, the phosphorothioate analogue exhibited stronger antiviral activity, the inhibition increased from 56 to 90%. The inhibitory effect of this analogue was antisense and sequence-specific. Northern blot analysis showed that the sequence-dependent mechanism of action appears to be the inhibition of mRNA transcription. We conclude that the coronavirus intergenic consensus sequence is a good target for an antisense oligonucleotide antiviral action. The properties of the phosphodiester oligonucleotide was improved after its complexation with cyclodextrins. The most important increase of the antiviral activity (90% inhibition) was obtained with only 7.5 microM oligonucleotide complexed to a cyclodextrin derivative, 6-deoxy-6-S-beta-D-galactopyranosyl-6-thio-cyclomalto-heptaose+ ++ in a molar ratio of 1:100. These studies suggest that the use of cyclodextrin derivatives as carrier for phosphodiester oligonucleotides delivery may be an effective method for increasing the therapeutic potential of these compounds in viral infections.

References1. Ho M, Chen ER, Hsu KH, et al. An epidemic of enterovirus 71 in Taiwan. N Engl J Med 1999; 341:929-35. 2. Lin TY, Twu SJ, Ho MS, Chang LY, Lee CY. Enterovirus 71 outbreaks, Taiwan: occurrence and recognition. Emerg Infect Dis 2003; 9: 291-3. 3. Hooi PS, Chua BH, Lee CS, Lam SK, Chua KB. Hand, foot and mouth disease: University Malaya medical center experience. Med J Malaysia

A fatal case of disseminated Scopulariopsis brevicaulis infection in an allogeneic stem cell transplant recipient is described. The patient was initially thought to have pulmonary aspergillosis, on the basis of clinical signs and antigenaemia, but Aspergillus was not isolated by culture. Scopulariopsis brevicaulis was subsequently isolated from skin and then from sputum and stool. Further investigation revealed that the infection had spread from a primary pulmonary site to the skin. A review of the literature underscores the difficulty of diagnosing infections caused by such emerging fungal pathogens and the poor outcome of immunocompromised patients with non-Aspergillus mould infections.

Emmonsia crescens is a saprophytic fungus that is distributed worldwide, causing diseases mostly in rodents. It has also been described, though rarely, as an etiologic agent of pulmonary pathology in humans, potentially leading to death. A case of pulmonary adiaspiromycosis is reported in a 30-year-old immunocompetent man. The patient presented with a history of several weeks of weakness, cough, fever, and weight loss of 10 kg. Clinical and radiographic findings showed pulmonary lesions consistent with tuberculosis or histoplasmosis, but no pathogen was found with classical microbiological procedures. The diagnosis of adiaspiromycosis due to Emmonsia crescens was initially made using molecular biology techniques. Histological observations subsequently confirmed the presence of adiaspores in granulomas. To our knowledge, this is the first case of adiaspiromycosis diagnosed by PCR and sequencing. The patient was treated with itraconazole and was seen at 1 month with symptomatic improvement. Here we will discuss this rare fungal infection and its difficult treatment and diagnosis. As represented in this case, molecular biology is a powerful method to optimize diagnostic tests and therefore improve the care of the infected patient. CASE REPORTA 30-year-old man was admitted to the Hospital of Metz on 21 May 2008 with a 2-week history of unproductive cough, fever at 39°C, progressive dyspnea, thoracic pain, generalized weakness, and weight loss of 10 kg in 1 month. The chest radiography revealed diffuse bilateral interstitial pneumonia with a micronodular pattern (Fig. 1). Computerized tomography of the chest (high resolution) showed disseminated pulmonary nodules (Fig. 2). The peripheral blood leukocyte count was 12 g/liter with 76% neutrophils and 14% lymphocytes. Bronchoalveolar lavage (BAL) showed 13,330 leukocytes/l with 1,333 lymphocytes (CD4/CD8 ϭ 1.25). The CRP was increased to 287 mg/liter, but no other abnormalities were found.Based on the clinical symptoms and radiological features, the main differential diagnosis was tuberculosis or histoplasmosis as the patient had traveled several months and years earlier in areas where these diseases were endemic. Other noninfectious hypotheses, such as lymphoma or sarcoidosis, were also suspected based on the clinical and biological features with an important alteration of the patient status associated with polyadenopathies. All of these hypotheses were then excluded by corresponding analyses.Diagnostic procedures. Direct examination and bacterial and mycological cultures of a BAL sample were negative. Serological tests for Histoplasma capsulatum were performed at the Pasteur Institute of Paris and revealed negative immunodiffusion. The BAL sample was sent to the Mycology Unit of the Nancy Hospital for further investigation.The BAL was microscopically reexamined with chlorazole black, and any fungal elements were visualized. Cultures of the sample on Sabouraud agar containing cycloheximide (0.5 mg/ ml) at 30°C and 37°C yielded no growth.Molecular biology studies. In or...