Direct amplification and species determination of microsporidian DNA from stool specimens

Trop Med Int Health. 1996 Jun;1(3):373-8. doi: 10.1046/j.1365-3156.1996.d01-51.x.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA Restriction Enzymes
  • DNA, Protozoan / analysis*
  • Feces / chemistry
  • Feces / parasitology*
  • Gene Amplification
  • Humans
  • Microsporida / genetics*
  • Microsporida / isolation & purification
  • Molecular Sequence Data
  • Polymerase Chain Reaction

Substances

  • DNA, Protozoan
  • DNA Restriction Enzymes