Document 0104
 DOCN  M9470104
 TI    Detection of microsporidia by indirect immunofluorescence antibody test
       using polyclonal and monoclonal antibodies.
 DT    9409
 AU    Aldras AM; Orenstein JM; Kotler DP; Shadduck JA; Didier ES; Department
       of Microbiology, Tulane University Regional Primate; Research Center,
       Covington, Louisiana 70433.
 SO    J Clin Microbiol. 1994 Mar;32(3):608-12. Unique Identifier : AIDSLINE
       MED/94253318
 AB    During a screening for monoclonal antibodies (MAbs) to the
       microsporidian Encephalitozoon hellem, three murine hybridoma cell lines
       producing strong enzyme-linked immunosorbent assay (ELISA) reactivities
       were cloned twice, were designated C12, E9, and E11, and were found to
       secrete MAbs to the immunoglobulin M isotype. On subsequent ELISAs, the
       three MAbs reacted most strongly to E. hellem, and they reacted somewhat
       less to Encephalitozoon cuniculi and least to Nosema corneum, two other
       microsporidian species. The MAbs produced values of absorbance against
       microsporidia that were at least three times greater than reactivities
       obtained with control hybridoma supernatants or with uninfected host
       cell proteins used as antigens. By Western blot immunodetection, the
       three MAbs detected three E. hellem antigens with relative molecular
       weights (M(r)s) of 62, 60, and 52 when assayed at the highest
       supernatant dilutions producing reactivity. At lower dilutions, the MAbs
       detected additional proteins with M(r)s of 55 and 53. By using indirect
       immunofluorescence antibody staining, the MAbs, as well as hyperimmune
       polyclonal murine antisera raised against E. cuniculi and E. hellem,
       were able to detect formalin-fixed, tissue culture-derived E. cuniculi
       and E. hellem and two other human microsporidia, Enterocytozoon bieneusi
       and Septata intestinalis, in formalin-fixed stool and urine,
       respectively. E. bieneusi, however, stained more intensely with the
       polyclonal antisera than with the MAbs. Neither the MAbs nor the
       hyperimmune murine polyclonal antibodies detected Cryptosporidium,
       Giardia, Trichomonas, or Isospora spp. At higher concentrations, the
       polyclonal antisera did stain N. corneum and yeast cells. The background
       staining could be absorbed with Candida albicans.(ABSTRACT TRUNCATED AT
       250 WORDS)
 DE    Animal  Antibodies, Monoclonal/DIAGNOSTIC USE  Antibodies,
       Protozoan/DIAGNOSTIC USE  AIDS-Related Opportunistic
       Infections/COMPLICATIONS/DIAGNOSIS/  PARASITOLOGY
       Encephalitozoon/*IMMUNOLOGY/ISOLATION & PURIF
       Encephalitozoonosis/COMPLICATIONS/*DIAGNOSIS/PARASITOLOGY  Evaluation
       Studies  *Fluorescent Antibody Technique  Human  Mice  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).