Immunodiagnosis of human fascioliasis by an enzyme-linked immunosorbent assay (ELISA) and a micro-ELISA.

Enzyme-linked immunosorbent assay (ELISA) and micro-ELISA were evaluated for their ability to detect anti-Fasciola hepatica antibodies in humans by using excretory-secretory antigen. The sensitivity of each method was 100%, but the specificity was 100% for ELISA and 97% for micro-ELISA. The micro-ELISA could be used as a screening assay and ELISA could be used as a confirmatory method for the serodiagnosis of human fascioliasis.

Diagnosis of Enterocytozoon bieneusi by PCR in stool samples eluted from filter paper disks.

We report a PCR-based assay for the detection of Enterocytozoon bieneusi. We extracted DNA from feces which had been applied to filter paper disks and evaluated four preserving solutions. Infected specimens were identified by electrophoresis of amplicons from concentrated formalin-fixed samples and unconcentrated fresh feces. Our findings demonstrate that this methodology is effective for sample collection, mailing, and diagnosis of this pathogen.

In situ hybridization: a molecular approach for the diagnosis of the microsporidian parasite Enterocytozoon bieneusi.

Microsporidia are emerging as opportunistic pathogens in patients with acquired immunodeficiency syndrome (AIDS). Enterocytozoon bieneusi is the most commonly reported microsporidium that is detected in gastrointestinal specimens. This report describes an in situ hybridization technique with a 30-base specific synthetic DNA probe for detection of E bieneusi by light microscopy. Formalin-fixed paraffin-embedded duodenal biopsy specimens from three patients with AIDS, chronic diarrhea, and E bieneusi infection confirmed by electron microscopy were used in this study. Light microscopic examination after colorimetric detection allowed the identification of different stages of the pathogen's life cycle in the cytoplasm of enterocytes. No cross-reactivity was noted between the probe and human DNA. Our study underscores the applicability of a synthetic-labeled oligonucleotide for the detection and identification of E bieneusi in clinical samples.

Estrategias clasicas para el dignostico de criptosporidiosis en pacientes con SIDA y diarrea cronica / Classic estrategies for diagnosis of cryptosporidiosis in patients with AIDS and chronic diarrhea

El protozoario Cryptosporidium sp. ha sido reconocido crecientemente en asociación con enteritis grave en paceintes con el síndrome de immunodeficiencia. Los individuos estudiados comprendieron 84 adultos com SIDA y diarrea crónica. En este trabajo se describen 14 pacientes con infección intestinal causada por Cryptosporidium sp. La media del recuerdo de CD4 en estos pacientes fue (300 células/mm3 (en 7 de los 14). El examen de aspirados duodenales y heces incluyó preparaciones de muestras concentradas coloreadas con Kinjoun, Dimetilsulfóxido y Auramina. Se realizaron videoesofagogastroduodenoscopías (VEDA) para inspeccionar visualmente la mucosa y obtener biopsias. La VEDA reveló duodeno granular en 10 pacientes y jaspeado en uno de ellos. Las biopsias duodenales fueron coloreadas con hematoxilina-eosina, Giemsa y Azur II. Los acambios histológicos incluyeron atrofia (3/14), duodenitis (2/14) a ambos (3/14). La microscopía eletrónica de transmisión fue usada para la identificación de estadíos de desarrollo de Cryptosporidium sp.

[Classic strategies for diagnosis of cryptosporidiosis in patients with AIDS and chronic diarrhea]. / Estrategias clásicas para el diagnóstico de criptosporidiosis en pacientes con SIDA y diarrea crónica.

Cryptosporidium sp., a protozoa organism, has been increasingly recognized in association with severe enteritis in patients with the Acquired Immunodeficiency Syndrome. The studied subjects included 84 adult patients with AIDS and chronic diarrhea. We describe 14 patients with intestinal infection caused by Cryptosporidium sp. The mean CD4 count in these patients was < or = 300 cells/mm3 (7 out of 14). Examination of duodenal aspirates and feces included dimethylsulfoxide, auramine and acid-fast preparation of concentrated samples. We carried out videoesophagogastroduodenoscopy (VEDA) to visually inspect the mucosa and obtain biopsy specimens. VEDA revealed granular duodenum in ten patients and jasper duodenum in one of them. Duodenal biopsy specimens were stained with hematoxylin-eosin, Giemsa and Azure II. Histologic changes included atrophy (3/14), duodenitis (2/14) or both (3/14). Transmission electron microscopy was used for the identification of developmental stages of Cryptosporidium sp.

[Morphological study of Enterocytozoon bineousi in patients with AIDS and chronic diarrhea]. / Estudio morfólogico de Enterocytozoon bieneusi en pacientes con SIDA y diarrhea crónica.

Microsporidia are protozoan parasites responsible for significant gastrointestinal disease in patients infected with the human immunodeficiency virus. We report the clinical features of three patients with chronic diarrhea and intestinal microsporidiosis caused by Enterocytozoon bieneusi. The average value for CD4 in these patients was < or = 50 cells/mm3. The spores were detected in smears from stool samples and duodenal aspirates stained with trichrome blue in all patients. Light microscopy of semi-thin plastic sections revealed parasites and spores in the enterocytes and were associated with villous atrophy (2 out of 3). Thin section-electron microscopy showed a variety of developmental stages of the microsporidio. Patients treated with Albendazole had an unsatisfactory clinical response to therapy. Enterocytozoon bieneusi infection may be an important cause of diarrhea in patients with AIDS in our country.

Estudio morfologico de Enterocytozoon bieneusi en pacientes con SIDA y diarrhea cronica / Morphological study of Enterocytozoon bineousi in patients with AIDS and chronic diarrhea

Microsporidia are protozoan parasites responsible for significant gastrointestinal disease in patients infected with the human inmunodeficiency virus. We reporte the clinical features of three patients with chronic diarrhea and intestinal microsporidiosis caused by Enterocytozoon bieneusi. The average value for CD4 in these patients was ó 50 cells/mm3. The spores were detected in smears from stool samples and duodenal aspirates stained with trichrome blue in all patiens. Light microscopy of semithin plastic sections revealed parasites and spores in the enterocytes and were associated with villous atrophy (2 out of 3). Thin section-electron microscopy showed a variety of developmental stages of the microsporidio. Patients treated with Albendazole had an unsatisfactory clinical response to therapy. Enterocytozzon bieneusi infection may be an important cause of diarrhea in patiens with AIDS in our country.

Detection of the microsporidian parasite Enterocytozoon bieneusi in specimens from patients with AIDS by PCR.

Microsporidia are protozoa parasites responsible for significant gastrointestinal disease in patients infected with human immunodeficiency virus. We evaluated a PCR assay of stool samples, duodenal aspirates, and biopsy specimens from patients with Enterocytozoon bieneusi infection. A 210-bp DNA fragment of the unique rRNA intergenic spacer could be amplified from all samples infected with E. bieneusi, but no amplification was seen by using DNA purified from samples with Septata intestinalis or other parasites and from negative control human cells. These results suggest that the PCR in stool samples may be a useful tool for the diagnosis of intestinal microsporidiosis in patients with AIDS.