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M9470104.TXT
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1994-07-02
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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).