期刊
INDIAN JOURNAL OF OPHTHALMOLOGY
卷 69, 期 9, 页码 2457-2460出版社
WOLTERS KLUWER MEDKNOW PUBLICATIONS
DOI: 10.4103/ijo.IJO_3685_20
关键词
Calcofluor white; corneal scraping; microbial keratitis; microscopic diagnosis; potassium hydroxide; trypan blue
资金
- Hyderabad Eye Research Foundation, Hyderabad, India
Trypan blue stain is highly efficacious in the diagnosis of fungal keratitis, showing comparable sensitivity and specificity to traditional KOH + CFW stain in the diagnosis of non-viral microbial keratitis. The study also found good agreement between observers in interpreting trypan blue-stained samples.
Purpose: To evaluate the efficacy of trypan blue in direct microscopic examination of corneal scrapings in the diagnosis of non-viral microbial keratitis. Methods: In a prospective, interventional, masked study, 82 consecutive patients were investigated. Direct microscopic examination of the corneal scrapings involved three smears stained with potassium hydroxide with calcofluor white (KOH + CFW), Gram stain (not analyzed), and trypan blue stain and culture for bacteria, fungus, and Acanthamoeba. While KOH + CFW stained slides were examined under a fluorescence microscope, trypan blue-stained slides were examined by two microbiologists (masked to KOH + CFW and culture results) under normal light microscopy. Thirty samples were reexamined for interobserver and intraobserver variability. Results: Out of 82 samples, fungal/fungus-like elements were seen in 48 (58.5%) in KOH + CFW and 38 (46.3%) in trypan blue stain. One microsporidial case, detected in KOH + CFW was negative in trypan blue stain (culture not done). Fungal growth was positive in 23 out of 81 (28.3%) cases cultured, single bacterial species in 18 (22.2%), Pythium insidiosum in three, mixed bacteria and fungus/Pythium in 7 (8.6%), Acanthamoeba in none and 30 (37.0%) samples were sterile. With culture as gold standard, the respective sensitivity (84.9%, 75.7%) and specificity (90.9%, 68.2%) of KOH + CFW stain and trypan blue stain were comparable (p = 0.16, P = 0.06). The interobserver linear weighted kappa score between the two microbiologists was 1.00 while it was 0.86 for intraobserver agreement. Conclusion: Trypan blue stain, an easily available dye to ophthalmologists, is highly efficacious in the diagnosis of fungal keratitis.
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