Title: Radhika Tandon, MD, DNB, FRCS, FRCOphth,
1COMPARISON OF TEAR COLLECTION TECHNIQUE FOR
DETECTION OF HSV DNA BY PCR
- Radhika Tandon, MD, DNB, FRCS, FRCOphth,
- Manoj Sharma, MD Gita Satpathy, MD
- Department of Cornea, Cataract Refractive
Surgery and Ocular Microbiology
Dr Rajendra Prasad Centre For Ophthalmic
Sciences, AIIMS
The authors have no financial interest in the
subject matter of this poster.
2 Review of Literature- Shedding of Herpes simplex
virus type 1 (HSV-1) DNA in active stromal
keratitis and / or endotheliitis, as detected by
polymerase chain reaction (PCR) analysis of tear
samples obtained by different methods
Authors (year) PCR type Subjects, proportion () Subjects, proportion () Sample Method Samples Collected per Subject
Authors (year) PCR type Active stromal keratitis Endotheliitis Sample Method Samples Collected per Subject
Yamamoto S et al (1994) Conventional PCR 2/6 (33.33) - Schirmer Strips 1
Kudo E et al (1996) Nested PCR 5/15 (33.33) - Schirmer Strips 3.3
Pramod NP et al (2000) (Conventional PCR 12/40 (30) - Schirmer Strips 2
Fukuda M et al (2003) Real-time PCR 8/14 (57) 0/3 Eye Rinse 1
Fukuda M et al (2008) Real-time PCR 13/22 (59.1) 0/3 Eye Rinse 1
Total no. of subjects with shedding of HSV-1
DNA/total no. of subjects assessed (percentage
of patients with shedding of HSV-1 DNA)
3AIM
- To compare PCR results in tear samples obtained
by two different methods - Schirmer's strip fire-polished micro capillary
tube - in patients with clinically diagnosed
- viral stromal keratitis and endotheliitis
4 Patients
- Inclusion Criteria
- Clinically diagnosed cases of active stromal
keratitis and endotheliitis - Exclusion criteria
- Pure epithelial keratitis with no stromal
involvement - History of oral acyclovir use within 1 month
5Study Design
- Study group 66 eyes( 59 patients)
- 52 unilateral 7 bilateral
- Control group 130 eyes of 90 patients
- Contralateral eye of 50 unilateral affected
patients - Both eyes of 40 normal volunteers
- Of 52 unilaterally affected, 2 patients had
contralateral phthisis bulbi, sample was not
taken from the phthisical eye
6Tear Collection Methods
- Tear samples of each patient were collected
- using Schirmers filter paper strip and fire
polished micro- capillary tubes in a randomized
sequence - from the anaesthetized inferior fornix
- and were subjected to PCR for detection of HSV
DNA
7RESULTS
Fig. 1 Distribution of tear samples from various
clinical categories PUK Peripheral ulcerative
keratitis A Active Q Quiescent P with
perforation V Viral
8Fig 2 PCR result in different clinical
categories with capillary Schirmers method
PUK Peripheral ulcerative keratitis A
Active Q Quiescent P with perforation V
Viral
9Figure 3 PCR result using Capillary and
Schirmer's tear sampling method
- PCR positivity in tear samples collected by
either method were not significantly different
(p0.23).
10Conclusion
- 20 cases of active Herpetic stromal keratits
and endotheliitis had positive tear PCR test
result - Tear samples collected by Schirmers strip and
fire- polished micro capillary tube are equally
effective for HSV DNA detection
11- Address for Correspondence
- Dr Radhika Tandon
- Professor of Ophthalmology
- radhika_tan_at_yahoo.com
- Dr. RP Centre for Ophthalmic Sciences,
- All India Institute of Medical Sciences
- (AIIMS)
- New Delhi, India