title: Evaluation of hematoxylin and eosin and special stains for the detection of acanthamoeba keratitis in penetrating keratoplasties.
abstract: OBJECTIVE
  To compare the efficacy of a battery of routine and special histologic stains for the detection of acanthamoeba keratitis.
  
  
  METHODS
  Observational study.
  
  
  METHODS
  Nine patients with culture-proven infectious keratitis whose clinical differential diagnosis included acanthamoeba and who had undergone penetrating keratoplasty were identified. Three cases each of culture-proven acanthamoeba, fungal, and herpes simplex keratitis were reviewed. Serial sections of the keratoplasty specimens were stained with hematoxylin and eosin, periodic acid-Schiff (PAS), Gomori methanamine silver (GMS), giemsa, Gram, calcofluor white, and acridine orange. Sections were reviewed in a random order and a masked fashion and classified as positive or negative for acanthamoeba, fungus, or herpes.
  
  
  RESULTS
  The correct diagnosis was made by examination of the hematoxylin and eosin stained slides in all cases. Correct diagnoses in decreasing order of frequency were made for slides stained with PAS, GMS, acridine orange, calcofluor white, giemsa, and Gram. There were false-positive diagnoses made only with calcofluor white and acridine orange stained slides because of staining of extracellular debris and other material.
  
  
  CONCLUSIONS
  When sections are examined by an experienced observer, hematoxylin and eosin is useful compared with calcofluor white, acridine orange, GMS, PAS, giemsa, and Gram stains for the detection of acanthamoeba keratitis.
authors.affiliation: Department of Ophthalmology, Emory University School of Medicine, Atlanta, Georgia, USA. ophtheg@emory.edu
authors.forename: Hans E
authors.initials: HE
authors.lastname: Grossniklaus
authors.forename: George O
authors.initials: GO
authors.lastname: Waring
authors.forename: Charlotte
authors.initials: C
authors.lastname: Akor
authors.forename: Amilcar A
authors.initials: AA
authors.lastname: Castellano-Sanchez
authors.forename: Kevin
authors.initials: K
authors.lastname: Bennett
mesh_terms: D015823:Acanthamoeba Keratitis / Q000175:diagnosis* / Q000601:surgery; D000165:Acridine Orange; D001557:Benzenesulfonates; D003315:Cornea / Q000382:microbiology / Q000469:parasitology / Q000821:virology; D004311:Double-Blind Method; D004801:Eosine Yellowish-(YS); D015821:Eye Infections, Fungal / Q000175:diagnosis / Q000601:surgery; D005189:False Positive Reactions; D005456:Fluorescent Dyes; D006416:Hematoxylin; D006801:Humans; D016849:Keratitis, Herpetic / Q000175:diagnosis / Q000601:surgery; D015948:Keratoplasty, Penetrating; D008856:Microscopy, Fluorescence; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D013194:Staining and Labeling / Q000379:methods
chemical_list: D001557:Benzenesulfonates; D005456:Fluorescent Dyes; C007061:C.I. Fluorescent Brightening Agent 28; D000165:Acridine Orange; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin