Molecular phylogenetic analysis of Enterobius vermicularis and development of an 18S ribosomal DNA-targeted diagnostic PCR.

We genetically characterized pinworms obtained from 37 children from different regions of Germany and established new species-specific molecular diagnostic tools. No ribosomal DNA diversity was found; the phylogenetic position of Enterobius vermicularis within the Oxyurida order and its close relationship to the Ascaridida and Spirurida orders was confirmed.

Genotyping of Paracoccidioides brasiliensis directly from paraffin embedded tissue.

Based on polymorphisms of the gp43 precursor gene, genotyping of Paracoccidioides brasiliensis was achieved in 6 out of 10 paraffin-embedded tissue samples by modifying a nested PCR procedure used in the diagnosis of the fungal infection. Nine of the samples originated in Brazil. Three sequences were identical to a previously published consensus sequence identifying the P.

Specific inhibitors of mitogen-activated protein kinase and PI3-K pathways impair immune responses by hemocytes of trematode intermediate host snails.

To characterize molecular mechanisms regulating snail cellular immune responses, the contributions of mitogen-activated protein kinases (MAPKs) and phosphatidylinositol 3-kinase (PI3-K) were examined in hemocytes of the trematode intermediate host snails Biomphalaria glabrata and Lymnaea stagnalis. Simultaneous measurement of phagocytosis/encapsulation and H2O2 production by hemocytes in the presence or absence of specific signal transduction inhibitors was used to assess the role of extracellular-signal regulated kinases 1 and 2 (ERK1/2), p38, JNK and PI3-K. Hemocyte spreading was significantly reduced in a dose-dependent manner by the ERK inhibitor, PD098059, and by wortmannin, a potent PI3-K inhibitor.

Coccidioidomycosis and blastomycosis: advances in molecular diagnosis.

Clinical isolates of Coccidioides spp. and Blastomyces dermatitidis can be identified by chemiluminescent DNA probes and PCR assays targeting multicopy genes. In fixed tissue samples, cells of the two fungi are specified by in situ hybridization and PCR assays targeting 18S rDNA but sequencing of the products is mandatory.

Comparison of autofluorescence and iodine staining for detection of Isospora belli in feces.

To evaluate the sensitivity of autofluorescence for detection of Isospora oocysts, wet preparations of 192 stool samples from patients with chronic diarrhea were examined by fluorescence microscopy and by light microscopy after iodine staining used for routine screening for ova and parasites. Silicon-chambered glass coverslips were used for fluorescence microscopy. Isospora oocysts were detected in 46 iodine-stained concentrated stool samples; 91 samples were positive by autofluorescence.

Comparison of fluorescence, antigen and PCR assays to detect Cryptosporidium parvum in fecal specimens.

To optimize routine screening for cryptosporidiosis, 198 stool samples from patients at risk and from calves were examined by enzyme immunoassay (EIA), a direct fluorescent-antibody (DFA) and a modified immunofluorescence assay. Ninety-nine samples were positive in at least one assay, whereas 99 were negative in all three assays. Sensitivity of antigen EIA and DFA were similar (94%, 95% CI: 88-98%, and 91%, 95% CI: 84-95%).