Identification of microsporidia in stool specimens by using PCR and restriction endonucleases.
We report the development of a PCR-based assay for the detection of microsporidia in clinical specimens. A single primer pair complementary to conserved sequences of the small-subunit rRNA enabled amplification of DNA from the four major microsporidian pathogens of humans: Encephalitozoon cuniculi, Encephalitozoon hellem, Enterocytozoon bieneusi, and Septata intestinalis. The extraction method allowed PCR amplification of E. bieneusi and S. intestinalis DNA from sodium hypochlorite-treated stool specimens. Differentiation of the microsporidian gastrointestinal pathogens E. bieneusi and S. intestinalis could be accomplished by restriction endonuclease digestion of PCR products using PstI and HaeIII.
Fedorko
D P
DP
Clinical Pathology Department, Warren G. Magnuson Clinical Center, National Institutes of Health, Bethesda, Maryland 20892, USA.
Nelson
N A
NA
Cartwright
C P
CP
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004262:DNA Restriction Enzymes; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification; D016819:Encephalitozoon / Q000235:genetics / Q000302:isolation & purification; D009670:Encephalitozoon cuniculi / Q000235:genetics / Q000302:isolation & purification; D005243:Feces / Q000469:parasitology*; D005767:Gastrointestinal Diseases / Q000469:parasitology; D006801:Humans; D016816:Microsporida / Q000235:genetics* / Q000302:isolation & purification*; D016881:Microsporidiosis / Q000469:parasitology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D016053:RNA, Protozoan / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics
D017931:DNA Primers; D016054:DNA, Protozoan; D016053:RNA, Protozoan; D012335:RNA, Ribosomal; D004262:DNA Restriction Enzymes