Direct amplification and species determination of microsporidian DNA from stool specimens.
Microsporidia are recognized as a major aetiological agent in chronic diarrhoea of immunocompromised patients. Their detection by light microscopy is hampered by the small size of the spores. A simple and rapid DNA extraction method has been developed for the detection of microsporidian DNA by PCR directly from stool specimens. It can be performed at room temperature in a 1.5-ml microcentrifuge tube format in less than 1 hour. The subsequent nested polymerase chain reaction permits the detection of 3-100 spores in a 0.1-g stool sample. The amplification products can be verified and the species Enterocytozoon bieneusi, Encephalitozoon cuniculi and Encephalitozoon (Septata) intestinalis distinguished by a simple restriction endonuclease digest.
Katzwinkel-Wladarsch
S
S
Department of Infectious Diseases and Tropical Medicine, University of Munich, Germany.
Lieb
M
M
Helse
W
W
Löscher
T
T
Rinder
H
H
D000818:Animals; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D016054:DNA, Protozoan / Q000032:analysis*; D005243:Feces / Q000737:chemistry / Q000469:parasitology*; D005784:Gene Amplification; D006801:Humans; D016816:Microsporida / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D016054:DNA, Protozoan; D004262:DNA Restriction Enzymes