       Document 0953
 DOCN  M9650953
 TI    Comparison of three staining methods for detecting microsporidia in
       fluids.
 DT    9505
 AU    Didier ES; Orenstein JM; Aldras A; Bertucci D; Rogers LB; Janney FA;
       Department of Microbiology, Tulane Regional Primate Research; Center,
       Covington, Louisiana 70433, USA. esdid@tpc.tulane.edu
 SO    J Clin Microbiol. 1995 Dec;33(12):3138-45. Unique Identifier : AIDSLINE
       MED/96156115
 AB    Calcofluor white 2MR, modified trichrome blue, and indirect
       immunofluorescent antibody (IFA) staining methods were evaluated and
       compared for detecting microsporidia in stool. Serial 10-fold dilutions
       of Encephalitozoon (Septata) intestinalis were prepared in three
       formalinized stool specimens or in Tris-buffered saline. Ten-microliter
       aliquots were smeared onto glass slides, fixed with methanol, stained,
       and read by at least three individuals. The results indicated that the
       calcofluor stain was the most sensitive method, required approximately
       15 min to perform, but did generate some false-positive results due to
       similarly staining small yeast cells. The modified trichrome blue stain
       was nearly as sensitive as the calcofluor stain and allowed for easier
       distinction between microsporidia and yeast cells. This stain, however,
       required approximately 60 min to perform. The IFA stain with polyclonal
       murine antiserum against E. intestinalis was the least sensitive of the
       methods and required approximately 130 min to perform. The lower limit
       of detection with the calcofluor and modified trichrome stains was a
       concentration of about 500 organisms in 10 microliters of stool to
       detect one microsporidian after viewing 50 fields at a final
       magnification of x1,000. Reliability was also addressed by use of 74
       stool, urine, and intestinal fluid specimens, 50 of which were confirmed
       for the presence of microsporidia by transmission electron microscopy
       (TEM). All TEM-positive specimens were detected by calcofluor and
       modified trichrome blue staining. Ten specimens were not detected by the
       IFA stain. An additional seven TEM-negative specimens were read positive
       for microsporidia with the calcofluor stain, and of these, five also
       were read positive with the modified trichrome blue stain. The resulting
       diagnostic paradigm was to screen specimens with the calcofluor stain
       and to confirm the results with the modified trichrome stain. IFA, which
       was less sensitive, may become useful for microsporidian species
       identification as specific antibodies become available.
 DE    Animal  AIDS-Related Opportunistic Infections/COMPLICATIONS/DIAGNOSIS/
       PARASITOLOGY  Benzenesulfonates  Body Fluids/PARASITOLOGY  Comparative
       Study  Diagnostic Errors  Encephalitozoon/ISOLATION & PURIF  Evaluation
       Studies  Feces/PARASITOLOGY  Fluorescent Antibody Technique,
       Indirect/STATISTICS & NUMER DATA  Fluorescent Dyes  Human  Microscopy,
       Electron  Microspora Infections/COMPLICATIONS/DIAGNOSIS/PARASITOLOGY
       Microsporida/*ISOLATION & PURIF  Reproducibility of Results  Sensitivity
       and Specificity  Staining/*METHODS/STATISTICS & NUMER DATA  Support,
       Non-U.S. Gov't  Support, U.S. Gov't, P.H.S.  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

