Detection of Herpes Simplex Virus Genome in Tear Fluid of Patients with Herpetic Keratouveitis and...
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Transcript of Detection of Herpes Simplex Virus Genome in Tear Fluid of Patients with Herpetic Keratouveitis and...
Detection of Herpes Simplex Virus Genome in Tear Fluid of Patients with Herpetic Keratouveitis and Endotheliitis
The authors have no financial interest in the subject matter of this poster
Department of Cornea, Cataract & Refractive Surgery and *Ocular Microbiology Dr Rajendra Prasad Centre For Ophthalmic Sciences, AIIMS
Radhika Tandon, MD, DNB, FRCS, FRCOphthDr. Manoj Sharma, MD,
Prof.Jeewan.S.Titiyal, MDDr Tushar Agarwal, MDDr Namrata Sharma, MDProf. Gita Satpathy, MD*
Aim
To evaluate the role of the polymerase chain reaction (PCR) for detecting Herpes virus DNA in tear samples of patients with herpetic keratouveitis and endotheliitis.
Patients
Inclusion criteria Clinically diagnosed Keratouveitis and/or endotheliitis
Exclusion criteria Oral acyclovir use within one month
Study Design Study group: 20 eyes (17 patients)
12 unilateral 3 bilaterally active 2 bilaterally affected with fellow eye quiescent*
Control group: 94 eyes of 54 patients Contra lateral eye of 14 unilateral active disease Both eyes of 40 normal volunteers
*59% (10/17) had definite history of recurrent disease
Laboratory Diagnosis Before starting treatment, tear samples were collected
with fire polished microcapillary tubes and subjected to PCR for HSV DNA detection
PCR Protocol
1.DNA extraction: commercial QI Amp DNA blood kit
2.Polymerase chain reaction
Primer-111 bp region of HSV 1 thymidine kinase gene (Hofgartner W T et al Clinical chemistry, 1999) Amplification- thermal cycler (Gene Amp PCR system 9700, applied biosystem, USA)
3.Electrophoreses- in 2% agarose gel
Treatment Tab acyclovir 400 mg (5 times/day) × 7 days followed by
Tab acyclovir 400 mg (BD) × 6 months Topical steroid (1% prednisolone acetate) Adjunct therapy was given as required
Topical antibiotic Topical mydriatic (2% homatropine) Topical lubricant (preservative free) Analgesics (if required)
Clinical type PCR TotalPositive Negative
Viral keratouveitis 2 9 11
Endotheliitis 2 7 9
Control 0 80 80
Fellow unaffected eyes 0 12 12
Fellow quiescent eye 0 2 2
Result
Four of twenty tear samples (20%) from patients yielded PCR positive for Herpes virus DNA, of whom 2 had acute endotheliitis and 2 had keratouveitis.
All four PCR positive cases had recurrent disease. No HSV DNA was detected in tear samples from both eyes of 40 normal
volunteers and fellow eye of unilateral active cases
At presentation 1 week 1 month (1a, 2a) (1b, 2b) (1c, 2c)
1a 1b 1c
2c2b2a
2 cases of Herpetic Keratouveitis who tested PCR positive in tear samples from the affected eye
1a 1b 1c
2c2b2a
A patient with bilateral Herpetic Endotheliitis who tested PCR positive in tear samples from both eyes
At presentation 1 week 3 weeks (1a, 2a) (1b, 2b) (1c, 2c)
Conclusion
20% cases of active keratouveitis and endotheliitis had positive tear sample PCR test result for HSV DNA
Herpes Virus DNA is detectable in tear samples of such patients and could prove useful in supporting the clinical diagnosis of these cases.
Address for Correspondence
Dr Radhika Tandon
Professor of Ophthalmology
Dr. RP Centre for Ophthalmic Sciences
All India Institute of Medical Sciences ( AIIMS)
New Delhi -110029
India