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William G. Stroop, Tsuey Ming Chen, James Chodosh, Thomas E. Kienzle, Janice L. Stroop, Jeng-Yang Ling, Dorothy A. Miles; PCR Assessment of HSV-1 Corneal Infection in Animals Treated with Rose Bengal and Lissamine Green B. Invest. Ophthalmol. Vis. Sci. 2000;41(8):2096-2102.
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purpose. In vivo, the ophthalmic dye rose bengal displays profound
antiviral effects against herpes simplex virus (HSV)-1, thus limiting
its utility in diagnosis of epithelial keratitis when used before viral
culture is performed. In contrast, lissamine green B does not possess
significant antiviral activity in vivo. To determine whether polymerase
chain reaction (PCR) could successfully detect HSV-1 DNA in ocular
samples that have been exposed to ophthalmic dyes, animal models were
used to observe the presence of infectious HSV-1 and viral DNA in eyes
treated with rose bengal or lissamine green B.
methods. Animals were bilaterally infected with HSV-1 strain H129, and at daily
intervals up to 16 days post infection (dpi) rose bengal or lissamine
green B was instilled in the left eyes. The right eyes were not treated
with dyes. Swabs of the dye-treated and untreated eyes were assayed by
PCR for viral infectivity by culture and the presence of DNA specific
for a fragment of the HSV-1 DNA polymerase gene.
results. A statistically equivalent number of samples from lissamine green
B–treated and untreated eyes were positive by both viral culture and
PCR. In contrast, rose bengal significantly decreased the infectious
virus present in ocular secretions. A total of 44% and 78% of the
rose bengal–treated and untreated eye samples, respectively, were
positive by culture from 1 through 16 dpi. PCR was more sensitive than
culture for detection of HSV-1 in rose bengal–treated eyes, in that
74% of rose bengal–treated samples were positive by PCR compared with
44% that were positive by culture during the 16-day period studied. It
was also noted that both rose bengal and lissamine green B treatments
slightly prolonged the period during which viral DNA was detectable in
ocular secretions by PCR, possibly because the singlet oxygen produced
by these photoreactive dyes compromised ocular cellular, humoral, and
nonspecific immune factors allowing viral DNA to persist for slightly
conclusions. PCR can successfully detect HSV-1 DNA in ocular samples that are
culture negative and contain rose bengal or lissamine green B.
Visualization of ocular epithelial defects with lissamine green B does
not interfere with detection of infectious virus or HSV-1
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