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Direct amplification and species determination of microsporidian DNA from stool specimens
Author(s) -
KatzwinkelWladarsch S.,
Lieb M.,
Heise W.,
Löscher T.,
Rinder H.
Publication year - 1996
Publication title -
tropical medicine and international health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.056
H-Index - 114
eISSN - 1365-3156
pISSN - 1360-2276
DOI - 10.1046/j.1365-3156.1996.d01-51.x
Subject(s) - microsporidia , enterocytozoon bieneusi , biology , microsporidiosis , polymerase chain reaction , microbiology and biotechnology , dna extraction , encephalitozoon cuniculi , spore , nested polymerase chain reaction , restriction enzyme , virology , dna , genetics , gene
Summary 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.