Polymerase chain reaction for diagnosis and species differentiation of microsporidia

Title:

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

Identifier:

https://cdk.lib.cas.cz/client/handle/uuid:3555262c-47ea-4ec7-94d7-3d4612842666
uuid:3555262c-47ea-4ec7-94d7-3d4612842666
urnnbn:urn:nbn:cz:aba007-000dp6

Subject:

microsporidia, Enterocytozoon bieneusi, Encephalitozoon, and polymerase chain reaction

Type:

model:internalpart and TEXT

Format:

print

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.

Language:

English

Rights:

http://creativecommons.org/licenses/by-nc-sa/4.0/
policy:public

Coverage:

140-148 and 62-70

Source:

Folia parasitologica | 1998 Volume:45 | Number:2

Harvested from:

CDK

Metadata only:

false