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

1999 ◽  
Vol 30 (1) ◽  
pp. 54-58 ◽  
Author(s):  
Jorge N Velasquez ◽  
Silvana Carnevale ◽  
Jorge H Labbe ◽  
Agustin Chertcoff ◽  
Marta G Cabrera ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Enterocytozoon Bieneusi ◽  
Molecular Approach ◽  
Microsporidian Parasite

scholarly journals Localization of Persistent Enterocytozoon bieneusiInfection in Normal Rhesus Macaques (Macaca mulatta) to the Hepatobiliary Tree

1998 ◽  
Vol 36 (8) ◽  
pp. 2336-2338 ◽  
Author(s):  
Keith G. Mansfield ◽  
Angela Carville ◽  
Daniel Hebert ◽  
Laura Chalifoux ◽  
Daniel Shvetz ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Macaca Mulatta ◽  
Rhesus Macaques ◽  
Clinical Signs ◽  
Enterocytozoon Bieneusi ◽  
Proximal Jejunum ◽  
Microsporidian Parasite ◽  
Immunodeficiency Virus ◽  
History Of

Enterocytozoon bieneusi is the most common microsporidian parasite recognized in human patients with AIDS. Recently, we identified a virtually identical organism causing a spontaneous infection associated with hepatobiliary and intestinal disease in simian immunodeficiency virus (SIV)-infected macaques. To examine the natural history of the infection, we examined captive rhesus macaques for E. bieneusi by PCR, in situ hybridization, and cytochemical techniques. PCR performed on fecal DNA detected enterocytozoon infection in 22 (16.7%) of 131 normal rhesus macaques (Macaca mulatta), compared to 18 (33.8%) of 53 rhesus macaques experimentally inoculated with SIV. In normal rhesus macaques, persistence of infection was demonstrated for up to 262 days and was usually not associated with clinical signs. In six of seven normal rhesus animals, E. bieneusi was detected by PCR in bile obtained through percutaneous cholecystocentesis but not by in situ hybridization performed on endoscopic biopsies of duodenum and proximal jejunum.

scholarly journals Prevalence of Enterocytozoon bieneusi in Swine: an 18-Month Survey at a Slaughterhouse in Massachusetts

2002 ◽  
Vol 68 (5) ◽  
pp. 2595-2599 ◽  
Author(s):  
Michael A. Buckholt ◽  
John H. Lee ◽  
Saul Tzipori
Keyword(s):  
In Situ Hybridization ◽  
Internal Transcribed Spacer ◽  
Enterocytozoon Bieneusi ◽  
Type D ◽  
Gnotobiotic Piglets

ABSTRACT Slaughterhouse pig samples were analyzed by PCR for Enterocytozoon bieneusi infection. Thirty-two percent were found to be positive, with rates being higher over the summer months. Three isolates from pigs were identical in their ribosomal internal transcribed spacer sequence to human E. bieneusi type D, two were identical to type F (from a pig), and nine were previously unreported. The viability of these spores was demonstrated by their ability to infect gnotobiotic piglets. The presence of the infection in liver was shown by in situ hybridization.

scholarly journals Transmission and Serial Propagation ofEnterocytozoon bieneusi from Humans and Rhesus Macaques in Gnotobiotic Piglets

1998 ◽  
Vol 66 (11) ◽  
pp. 5515-5519 ◽  
Author(s):  
Ivanela Kondova ◽  
Keith Mansfield ◽  
Michael A. Buckholt ◽  
Barry Stein ◽  
Giovanni Widmer ◽  
...  
Keyword(s):  
Animal Model ◽  
In Situ Hybridization ◽  
Chronic Diarrhea ◽  
Rhesus Macaques ◽  
Scientific Progress ◽  
Enterocytozoon Bieneusi ◽  
Oral Challenge ◽  
Infected Piglet ◽  
Gnotobiotic Piglets

ABSTRACT For over a decade Enterocytozoon bieneusi infections in people with AIDS have been linked with chronic diarrhea and wasting. The slow scientific progress in treating these infections is attributed to the inability of investigators to cultivate the parasite, which has also precluded evaluation of effective therapies. We report here successful serial transmissions of E. bieneusi from patients with AIDS and from macaques with AIDS to immunosuppressed gnotobiotic piglets. One infected piglet was still excreting spores at necropsy 50 days after an oral challenge. Spores in feces were detected microscopically by trichrome stain and by PCR and within enterocytes by in situ hybridization and immunohistochemistry. E. bieneusiinfection induced no symptoms. The development of an animal model forE. bieneusi will open up new opportunities for investigating this parasite.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

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