Indian Journal of Ophthalmology

: 2000  |  Volume : 48  |  Issue : 2  |  Page : 140--1

Herpes simplex virus DNA in the lens one year after an episode of retinitis

LK Therese, K Priya, SK Rao, J Biswas, HN Madhavan 
 Vision Research Foundation, Chennai, India

Correspondence Address:
L K Therese
Vision Research Foundation, Chennai

How to cite this article:
Therese L K, Priya K, Rao S K, Biswas J, Madhavan H N. Herpes simplex virus DNA in the lens one year after an episode of retinitis.Indian J Ophthalmol 2000;48:140-1

How to cite this URL:
Therese L K, Priya K, Rao S K, Biswas J, Madhavan H N. Herpes simplex virus DNA in the lens one year after an episode of retinitis. Indian J Ophthalmol [serial online] 2000 [cited 2022 Nov 28 ];48:140-1
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Full Text

Herpes simplex virus (HSV) is established as one of the causative agents of acute retinal necrosis (ARN) syndrome. We have earlier reported the first instance of HSV-1 isolation from intraocular fluid containing lens aspirate three-and-a-half months after complete regression of the disease in a patient of ARN.[1]

 Case report

On January 8, 1997, a 33-year-old man presented with clinical features of acute retinal necrosis (ARN) in the right eye and underwent investigations for viral aetiology. Immunofluorescence on aqueous humor (AH) collected, as diagnostic aspirate did not show the presence of Herpes simplex virus (HSV), Cytomegalovirus (CMV) or Varicella zoster virus (VZV) antigens. Virus isolation was negative for all the three viruses: for VZV and CMV in diploid cell line MRC-5, and for HSV in Vero cell line obtained from the National Facility for Animal Tissue Cell Culture (NFATCC), Pune, India. Antibodies were not detected in venous blood sample against HSV, CMV and VZV by ELISA and against human immunodeficiency virus (HIV) 1 and 2 by Tridot (J. Mitra, Chennai) and Immunocomb (OSB agencies, New Delhi).

Polymerase chain reaction (PCR) using primers coding for DNA polymerase gene as described by Cunningham et al[2] and Lakeman et al[3] with the primer sequences 5' - ATC AAC TTC GAC TGG CCC TT - 3' & 5' - CCG TAC ATG TCG ATG TTC AC - 3' (custom synthesized by Bangalore Genei Private Ltd, India) generating a 179 bp product was standardised in our laboratory. This PCR was found to be specific (determined by using related bacteria and viruses), and sensitive enough (determined by using 10-fold dilutions of DNA from HSV-1 [ATCC 733-VR] & HSV-2 [SP 753167]) to detect 0.5 fg of HSV-land 0.2 fg of HSV-2 DNA. We applied this technique in the AH specimen and detected the presence of HSV-DNA Figure.

The patient was treated for HSV infection based on the clinical presentation. Significant clinical improvement occurred with intravenous acyclovir, oral prednisolone and topical 0.1% betamethasone. Since persistent vitreous haze interfered with retinal photocoagulation, vitrectomy and endolaser photocoagulation were performed on January 20, 1997. Vitreous fluid (VF) was collected at the beginning of vitrectomy. Immunofluorescence on VF did not show the presence of HSV, CMV, and VZV antigens. Virus isolation was negative for all the three viruses: for VZV, CMV and HSV. A second convalescent blood sample was collected 17 days after the first. ELISA on acute serum sample was negative for antibodies against HSV, CMV and VZV while the convalescent sample showed the presence of anti-HSV IgG at 1:100 dilution and anti-CMV IgG at 1:40 dilution. Antibodies against VZV were not detected in the convalescent sample.

PCR of the VF was negative for HSV-DNA Figure. This was due to the presence of inhibitors of PCR for HSV-DNA in the VF because amplification of HSV-DNA did not occur when PCR was done on VF after spiking it with HSV-DNA. The presence of inhibitors in VF was reconfirmed by performing PCR on freshly extracted DNA from the VF spiked with HSV-DNA.

A year later, posterior subcapsular cataract formation resulted in decreased visual acuity. Phacoemulsification and intraocular lens implantation were performed in February 1998. During surgery, lens cortical material was obtained for microbiological analysis before phacoemulsification. Immunofluorescence on the direct smear and culture did not reveal the presence of HSV in the cortical lens material, but PCR was positive for HSV-DNA in the same Figure.


Aetiological diagnosis of HSV infection in this case of retinitis was supported by detection of HSV-DNA in AH by PCR, minimal rise of anti-HSV antibody in the convalescent serum and therapeutic response to acyclovir. Our result showed that PCR on AH was a useful tool to detect HSV in ARN. PCR for HSV-DNA on VF may be negative due to the presence of inhibitors. Since anterior chamber tap is a less invasive and hazardous procedure than the vitreous aspiration and can also be performed in the office, AH so obtained could be subjected to PCR in cases of viral retinitis.[4] Detection of HSV-DNA in the cortical lens material one year following ARN indicates persistence of HSV within the eye, possibly in the lens, even after inflammation has subsided completely in ARN. Contamination of the lens cortex by HSV-DNA from AH during cataract surgery is also a possibility, but its detection was indicative of the presence of the latent virus in the eye. Such latent HSV may have a potential role in recurrence due to reactivation.[5]


1Biswas J, Madhavan HN, Badrinath SS. Demonstration of herpes simplex virus from lens aspirate in healed acute retinal necrosis syndrome. Br J Ophthalmol 1997;81:802-3.
2Cunningham ET, Short GA, Irvine AR, Duker JS, Margolis TP. Acquired immunodeficiency syndrome-associated herpes simplex retinitis. Clinical description and use of a polymerase chain reaction-based assay as a diagnostic tool. Arch Ophthalmol 1996;114:834-40.
3Lakeman FD, Whitley RJ. Diagnosis of Herpes simplex encephalitis: application of polymerase chain reaction to cerebrospinal fluid from brain-biopsied patients and correlation with disease. J Infect Dis 1995;171:857-63.
4Fox GM, Crouse CA, Chuang EL, Pflugfelder SC, Cleary TJ, Nelson SJ, et al. Detection of Herpesvirus DNA in vitreous and aqueous specimens by the polymerase chain reaction. Arch Ophthalmol 1991;109:266-71.
5Rahhal FM, Siegel LM, Russak V, Wiley CA, Tedder DG, Wienberg A, et al. Clinicopathologic correlations in acute retinal necrosis caused by Herpes simplex virus type 2. Arch Ophthalmol 1996;114:1416-19.