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2. Polymerase chain reaction for diagnosis and species differentiation of microsporidia

Creator:
Franzen, Caspar, Muller, Andreas, Hartmann, Pia, Hegener, Petra, Schrappe, Matthias, Diehl, Volker, Fatkenheuer, Gerd, and Salzberger, Bernd
Format:
print
Type:
model:internalpart and TEXT
Subject:
microsporidia, Enterocytozoon bieneusi, Encephalitozoon, and polymerase chain reaction
Language:
English
Description:
Polymerase chain reaction (PCR) techniques have been developed for the detection of microsporidian DNA in different biological samples. We used sequence data of the rRNA gene for the identification of Enterocytozoon bieneusi, Encephalitozoon intestinalis, E. cuniculi, and E. hellem in different biological samples of HIV-infected patients by PCR, Southern blot hybridization, restriction endonuclease digestion analysis, cloning, and comparative genetic sequencing. One primer pair was used for amplification of the entire small subunit (SSU)-rRNA gene of E. bieneusi, E, intestinalis, and E. hellem from samples with electron microscopy confirmed infection. The amplified 1.2 kb SSU-rRNA gene fragments were ligated into a pMOSBlue T-vector, transfected into pMOSS/ме competent cells, and were used as positive controls. Several primer pairs and hybridization probes were used to amplify and identify microsporidian DNA from different samples. Light microscopical examination of samples was performed in all patients and transmission electron microscopy was done on a subset of patient samples. DNA products were obtained from all samples with confirmed microsporidial infections. The identity of the DNA fragments was determined by Southern blot hybridization or by restriction endonuclease digestion analysis or by DNA sequencing. The results show that PCR is a reliable and sensitive indicator for the presence of microsporidian DNA in different biological samples of HIV-infected patients. PCR can be used further for species differentiation of microsporidia, even between species which cannot be differentiated by light and/or electron microscopy.
Rights:
http://creativecommons.org/licenses/by-nc-sa/4.0/ and policy:public

3. Two hemocyte populations in Triatoma infestans: ultrastructural and lectin-binding characterization

Creator:
Hypša, Václav and Grubhoffer, Libor
Format:
print
Type:
model:internalpart and TEXT
Language:
English
Description:
Two distinct hemocyte populations are determined in the hemolymph of the triatomine bug Triatoma infestans Klug, oenocytoids and plasmatocytes, and their independent origin from separate stem cells is shown. Both hemocyte populations differ considerably in their morphology, ultrastructure and lectin-binding properties. While oenocytoids are quite uniform with easily definable cells which do not to bind any assayed lectin, the plasmatocytes are a very polymorphic population possessing several morphological types and displaying a positive reactivity with lectins.
Rights:
http://creativecommons.org/licenses/by-nc-sa/4.0/ and policy:public